PT1682606E - Anion-binding polymers and uses thereof - Google Patents

Abstract

Anion-binding polymers are described. The anion-binding polymers in some cases are low swelling anion-binding polymers. In some cases, the anion-binding polymers have a pore volume distribution such that a fraction of the polymer is not available for non-interacting solutes above a certain percentage of the MW of the target ion for the polymer. In some cases, the anion-binding polymers are characterized by low ion-binding interference, where the interference is measured in, for example, a gastrointestinal simulant, relative to non-interfering buffer. Pharmaceutical composition, methods of use, and kits are also described.

Description

DESCRIPTION " POLYMERS CONNECTING TO ANIONS AND THEIR USES "

BACKGROUND OF THE INVENTION

Ion selective sorbents have been used in human therapy to correct electrolyte disturbances in conditions such as hyperphosphatemia, hyperoxaluria, hypercalcemia and hyperkalemia. Hyperphosphataemia occurs in patients with renal insufficiency, whose kidneys have ceased to excrete enough phosphate ions to compensate for exogenous uptake of phosphate from the diet. This pathology leads to elevated serum phosphate concentrations and to a high calcium phosphate product. Although the etiology is not fully demonstrated, the high calcium-phosphate product has been considered responsible for soft tissue calcification and cardiovascular disease. Cardiovascular disease is the cause of death in almost half of patients undergoing dialysis.

Aluminum, calcium and, most recently, lanthanum salts have been prescribed to control the absorption of phosphate ions in the gastrointestinal tract (GI) and to restore systemic phosphate levels to normal. However, these salts release soluble calcium and aluminum cations into the GI tract, which are then partially absorbed into the bloodstream. Absorption of aluminum can lead to serious side effects such as bone disease and aluminum dementia; elevated calcium uptake leads to hypercalcaemia and puts patients at risk of coronary calcification. 1

The non-metal phosphate binders, such as the strong basic ion exchange materials, Dowex resins and Cholestyramine, have been suggested for use as phosphate binders. However, its low binding capacity requires a high dosage that is not well tolerated by patients.

Functional amine polymers have been described as phosphate or oxalate binders. For example see documents 5985938; 5,980,881; 6180094; 6423754; and PCT publication WO 95/05184. Renagel, a crosslinked polyallylamine resin, is a phosphate sequestering material which is marketed as a binder without metal. Renagel phosphate binding in vitro is approximately 6 mmol / g in water and 2.5 mmol / g when measured in 100 mM sodium chloride and 20 mM phosphate at neutral pH. The recommended dosage for the target patient population is typically from 5 g / day to 15 g / day to maintain the phosphate concentration below 6 mg / dL. Renagel published Phase I clinical trials in healthy volunteers indicate that 15 g of Renagel decreases urinary phosphate excretion from a reference value of 25 mmol to 17 mmol and the difference is excreted in faeces in the form of phosphate free and polymer bound. From these data the range of in vivo capacity can be established at 0.5-1 mmol / g, which is much lower than the in vitro capacity of 6 mmol / g measured in physiological saline. Considering only the in vitro binding capacity of the measured Renagel in saline, a 15 g dosage of the phosphate binder would bind more than the total phosphorus content present in the average American diet, i.e. 37 mmol / day. The discrepancy between in vitro binding ability and documented low binding capacity in vivo has a negative impact on the therapeutic benefit of the drug since more resin is required to bring serum phosphate into a safe range.

This loss of capacity of the ion exchange resins is not restricted to Renagel when used in the complex medium of the GI tract. While generally safe from a toxicological perspective, the high dose and inconvenience associated with ingesting amounts of the order of several grams of resin argues in favor of the need to improve the capacity of the resin. As an example, even in safety studies described for the Renagel binder, patients experienced gastrointestinal discomfort at doses as low as 1.2-2.0 g / day over an 8-week treatment period.

Patients receiving 5.4 g of Renagel / day discontinued treatment due to adverse events such as GI discomfort in 8.9% of cases (Slatapolsky, et al., Kidney Int 55: 299-307, 1999; Chertow et al. Nephrol Dial Transplant 14: 2907-2914, 1999). Thus, an improvement in in vivo binding capacity which results in lower, better tolerated dosages would be a welcome improvement in resin based therapies.

As a result of these considerations there remains a great need for high capacity, safe binders that selectively eliminate ions from the body with a lower dosage of the drug and a better patient compliance profile. Adherence by the patient is currently recognized as one of the key factors for patients to comply with K / DOQI recommendations: increasing the dose implies that patients have to take ten 800 mg pills per day and more. 3

Renagel pills appear in the form of tablets to be ingested and are administered with a minimal amount of fluid, which further causes difficulties for ESRD patients who are under fluid restriction. An easier-to-take pharmaceutical formulation would be desirable: in particular chewable tablets are becoming more popular among the pediatric and geriatric population and in treatments requiring a large pill effort: chewable tablets allow higher potency pills and they ultimately reduce the number of pills per meal. Since the active substance present in a chewable tablet is mainly dispersed by the chewing and saliva effects before being swallowed, the requirements in terms of tablet form and weight are much less stringent than those imposed on tablets to be swallowed. However, to date, it has not been possible to formulate a hydrogel, such as Renagel, in a chewable tablet due to the high dilation characteristics of that polymer: Renagel generally dilates very rapidly to about 10 times its weight in a solution isotonic. This has two very undesirable consequences: firstly, while in the mouth, the polymer will dilate causing a very unpleasant sensation (dry mouth, feeling of suffocation); Second, even if the patient overcomes the sensation in the mouth, administering a dilated gel into the esophagus can be dangerous. Furthermore, it is also well known that, when administered over a range of several grams, the extremely swellable gels cause side effects, such as swelling, intestinal constipation or diarrhea. US 5338532 describes star-dense conjugates comprising a branched dendritic molecule. JP 2003 155429 A discloses ink jet compositions comprising a " mycelial forming component " with amine functionality. US 5532092 describes dendritic macromolecules comprising a nucleus and branching from the nucleus, wherein the dendritic macromolecule comprises amine groups.

SUMMARY OF THE INVENTIVE CONCEPT

In one aspect, the inventive concept provides anion-binding polymers. In some embodiments, the inventive concept provides an anion-binding polymer wherein the polymer binds to a target anion (eg, phosphate or oxalate), and wherein the polymer is characterized by at least two of the following particularities : a) a dilation rate of less than about 5; b) a pore volume distribution of the gel measured in a physiological medium characterized by a fraction of said pore volume, accessible to non-interacting solutes of molecular weight greater than about twice the MW of the target anion, less than about 20% by weight gel; and c) an ion-binding interference to the target anion of less than about 60% when measured in a gastrointestinal mock relative to a non-interfering buffer. In some embodiments, the rate of dilation is less than about 4, or less than about 3, or less than about 2.8, or less than about 2.7, or less than about of 2.6, or less than about 2.5. In some embodiments, the polymer binds bile acids or citrate having a capacity of less than about 2 mmol / g, or less than about 1 mmol / g, or less than about 0.5 mmol / g, or less than about 0.3 mm / g, or less 5 than about 0.1 mm / g. In some embodiments, the rate of dilation is measured in isotonic solution and neutral pH. In some embodiments, the polymer comprises amine monomers. In some embodiments, the amine monomers are selected from the group consisting of allylamine, vinylamine, ethyleneimine, 1,3-diaminopropane, and Ν, Ν, Ν ', N'-tetrakis (3-aminopropyl) -1,4- diaminobutane, 1,2,3,4-tetraaminobutane, Formula 1 and Formula 2, wherein Formula 1 and Formula 2 are the following structures:

NHa

In some embodiments, the inventive concept provides an anion-binding polymer containing crosslinked polyamines, wherein the polymer is obtained by reverse suspension, and wherein the rate of expansion of the polymer is less than 5.

In some embodiments, the inventive concept provides a phosphate-binding polymer wherein the polymer is characterized by at least one of the following particularities: a) a dilation rate of less than about 5, preferably less than which is about 2.5; b) a pore volume distribution of the gel, measured in a physiological medium characterized by a fraction of said pore volume, accessible to non-interacting solutes of molecular weight greater than about 200, less than about 20% by weight of gel; and c) an interference at ion-to-phosphate binding of less than about 60% when measured in a gastrointestinal mock, relative to a non-interfering buffer. In some embodiments, the rate of dilation is less than about 2.8, or less than about 2.7, or less than about 2.6. In some embodiments, the polymer binds bile acids or citrate having a capacity of less than about 2 mmol / g, or less than about 1 mmol / g, or less than about 0.5 mmol / g, or less than about 0.3 mm / g, or less than about 0.1 mm / g. In some embodiments, the rate of dilation is measured in isotonic solution and neutral pH.

In some embodiments, the inventive concept provides a phosphate-binding polymer wherein the polymer is characterized by a dilation rate of less than about 5, preferably less than about 2.8, or less than about 2.7, or less than about 2.6, most preferably less than about 2.5, wherein this rate is measured in isotonic solution and at neutral pH. In embodiments, the polymer has a mean in vivo phosphate binding capacity of greater than about 0.5 mol / g. In embodiments, the polymer is a polyamine polymer, and the chloride content of the polymer is less than about 35 mol% of the amine group content.

In some embodiments, the inventive concept provides an anion-binding polymer in which the polymer binds a target anion (eg, phosphate or oxalate), and wherein the polymer is characterized by at least two of the following particularities: a) a dilation rate of less than about 5; b) a pore volume distribution of the gel measured in a physiological medium characterized by a fraction of said pore volume, accessible to non-interacting solutes of molecular weight greater than about twice the MW of the target anion, less than about of 20% by weight of the gel; and c) an ion-binding interference to the target anion of less than about 60% when measured in a gastrointestinal mock, relative to a non-interfering buffer, wherein the polymer contains one or more amine monomers and one or more crosslinkers, and that the polymer is produced by a process in which the amine is present in the solvent prior to crosslinking in an amine: solvent ratio of from about 3: 1 to about 1: 3 and the total content of crosslinking agents added to the reaction mixture is such that that the average number of bonds to the amine (NC) monomers is between about 2.05 and about 6, or between about 2.2 and about 4.5. In some embodiments, the polymer is further produced by a process wherein the target anion is present during the crosslinking reaction, e.g. g): a) adding the amine monomer as a free base and adding the target anion to its acid form; b) adding a crosslinker, c) carrying out the crosslinking reaction; and d) eliminating the target ion by washing.

In some embodiments, the inventive concept provides an anion-binding polymer in which the polymer binds a target anion (eg, phosphate or oxalate), and wherein the polymer is characterized by at least two of the following particularities: a) a dilation rate of less than about 5; b) a pore volume distribution of the gel measured in a physiological medium, characterized by a fraction of said pore volume, accessible to non-interacting solutes of molecular weight greater than about twice the MW of the target anion, less than about of 20% by weight of the gel; and c) an ion-binding interference to the target anion of less than about 60% when measured in a gastrointestinal mock, relative to a non-interfering buffer, wherein the polymer contains one or more amine monomers and one or more crosslinkers, and that the polymer is produced by a process comprising: a) preparing a soluble prepolymer by adding the entire amine monomer component and then adding a fraction of the crosslinker continuously to form a syrup; b) emulsifying the syrup in oil; and c) adding the remaining crosslinking moiety to form crosslinked beads.

In some embodiments, the inventive concept provides an anion-binding polymer wherein the polymer binds a target anion (eg, phosphate or oxalate), and wherein the polymer is characterized by at least two of the following particularities: a ) a dilation rate of less than about 5; b) a pore volume distribution of the gel measured in a physiological medium characterized by a fraction of said pore volume, accessible to non-interacting solutes of molecular weight greater than about twice the MW of the target anion, less than about 20% by weight gel; and c) an ion-binding interference to the target anion of less than about 60% when measured in a gastrointestinal mock, relative to a non-interfering buffer, wherein the polymer contains one or more amine monomers and one or more crosslinkers, and that the polymer is produced by a process comprising: a) performing a first reaction between an amine monomer and a crosslinker to form a gel; followed by b) reacting the gel with an aminoalkyl halide, wherein the aminoalkyl groups are chemically linked to the gel by substituting the halide for the amine functional gels.

In some embodiments, the inventive concept provides a phosphate-binding polymer containing one or more amine monomers and one or more crosslinkers wherein the polymer is produced by a process wherein the total content of crosslinkers added to the reaction mixture is so the average number of bonds to the amine monomers being between 2.2 and 4.5.

In some such embodiments, the amine monomer is selected from the group consisting of 1,3-diaminopropane and Ν, Ν, Ν ', N'-tetrakis (3-aminopropyl) -1,4-diaminobutane, and wherein the crosslinker is selected from the group consisting of 1,3-dichloropropane and epichlorohydrin. In embodiments, the invention provides an ion-binding polymer comprising epichlorohydrin cross-linked Ν, Ν, Ν ', N'-tetrakis (3-aminopropyl) -1,4-diaminobutane wherein the polymer is produced by a the ratio of the initial concentration of Ν, Ν, Ν ', N'-tetrakis (3-aminopropyl) -1,4-diaminobutane to water is about 1: 3 to about 4: 1, or about 1.5: 1 to about 4: 1.

In some embodiments, the invention provides a phosphate-binding polymer containing Ν, Ν, Ν ', N'-tetrakis (3-aminopropyl) -1,4-diaminobutane and cross-linking epichlorohydrin monomers, wherein the polymer is produced by a process in which the total cross-linking epichlorohydrin added to the reaction mixture is about 200% to about 300 mol%, or about 230 to about 270 mol%, or about 250 mol% of the total Ν , Ν, Ν ', N'-tetrakis (3-aminopropyl) -1,4-diaminobutane. In some of these embodiments, the polymer 10 is produced by a process in which the ratio of the monomers to the water in the initial reaction mixture is about 3: 1 to about 1: 1, or about 1.73. In some embodiments, the polymer is in the form of spherical beads.

In some embodiments, the inventive concept provides a phosphate-binding polymer comprising polyallylamine monomers and cross-linking epichlorohydrin, wherein the polymer is produced by initially dissolving the polyallylamine monomers in water in a monomer: water ratio of about 3: 1 to about 1: 3. In some of these embodiments, the total crosslinking epichlorohydrin added to the reaction mixture is about 10 mol% of the total polyallylamine content.

In some embodiments, the inventive concept provides a phosphate-binding polymer comprising a prepolymer comprising 1,3-diamino propane and cross-linking 1,3-dichloropropane in a 1: 1 molar ratio, wherein the prepolymer is still crosslinked with the cross-linker epichlorohydrin, and wherein the total cross-linking epichlorohydrin added to the reaction mixture is about 200 mol% of the total prepolymer, and wherein the prepolymer: water ratio in the reaction mixture is about 1.1: 1 to about 1.7: 1. The inventive concept further provides compositions containing any of the foregoing polymers wherein the polymer is in particulate form and wherein the polymer particles are enclosed in an outer coating.

In another aspect, the invention provides pharmaceutical compositions. In one embodiment, the pharmaceutical composition contains a polymer of the invention and a pharmaceutically acceptable excipient. In some embodiments, the composition is a liquid formulation in which the polymer is dispersed in a liquid carrier of water and suitable excipients. In some embodiments, the inventive concept provides a pharmaceutical composition comprising an anion-binding polymer that binds a target anion, and one or more suitable pharmaceutical excipients, wherein the composition is in the form of a chewable or disintegrating tablet in mouth, and wherein the polymer has a rate of dilation as it passes through the oral cavity and the esophagus of less than about 5, or less than about 2.8, or less than about 2.7 or less of the which is about 2.6, or preferably less than about 2.5. In some embodiments the chewable tablet contains polymer wherein the polymer has a transition temperature greater than about 50Â ° C.

In some embodiments the chewable tablet contains a pharmaceutical excipient selected from the group consisting of sucrose, mannitol, xylitol, maltodextrin, fructose, sorbitol and combinations thereof, and is produced by a process wherein the polymer is pre-formulated with the excipient for forming a solid solution. In some embodiments the target anion of the polymer is phosphate. In some embodiments the polymer binds a target ion in vivo with a binding activity greater than 0.5 mmol / g. In some embodiments the anion-binding polymer is more than about 50% by weight of the tablet. In some embodiments, the tablet is cylindrically shaped with a diameter of about 22 mm and a height of about 4 mm and the anion-binding polymer constitutes more than about 1.6 g of the total weight of the tablet. In some of the chewable tablets of the inventive concept, the excipients are selected from the group consisting of sweeteners, binders, lubricants and disintegrants. Optionally, the polymer is present as particles with less than about 40 μm of average diameter. In some of these embodiments, the sweetener is selected from the group consisting of sucrose, mannitol, xylitol, maltodextrin, fructose and sorbitol, and combinations thereof.

In another aspect, the inventive concept provides a method for measuring interference at ion-binding to an ion-binding polymer that binds a target ion: a) by adding the ion-binding polymer to a non-interfering buffer containing the target ion and measuring the ability of the polymer to bind to the target ion; b) artificially digesting a standard meal with mammalian GI enzymes and / or aspirating chyme from the upper gastrointestinal tract of mammals that ingested said standard meal; wherein the standard meal contains the target ion; c) adding the ion-binding polymer and measuring the binding capacity of the target ion from the decrease in the concentration of the target ion after and prior to the addition of the target ion; and (d) by calculating the degree of interference in the binding as the fractional decrease in targeting ability expressed as a percentage between the measurement of binding in a non-interfering buffer and in the digested meal or liquids harvested ex vivo at the same concentration of the ion in equilibrium.

In yet another aspect, the inventive concept provides a method for selecting an ion-binding polymer, said polymer comprising monomer and crosslinker, wherein said polymer has at least one of the features: a) a rate of expansion less than about 135; b) a pore volume distribution of the gel measured in a physiological medium characterized by a fraction of said pore volume, accessible to non-interacting solutes of molecular weight greater than about twice the MW of the target anion, less than about of 20% by weight of the gel; and c) an ion-binding interference to the target anion of less than about 60% when measured in a gastrointestinal mock relative to a non-interfering buffer comprising the steps of: i) varying the following composition and process variables: 1) the ratio of crosslinker to monomer; 2) the ratio of (monomer + crosslinker) to the solvent in the reaction medium; 3) the net polymer load at physiological pH and tonicity; and / or 4) the hydrophilic / hydrophobic balance of the polymer backbone; ii) evaluate the dilatability, porosity and interference in the ion binding of the resulting polymer; and iii) selecting a polymer having at least one of said features. In another aspect, the inventive concept provides a method for improving the therapeutic and / or aptitude for administration and / or pharmaceutical properties of a polyamine polymer comprising at least one of the following steps: a) crosslinking said polymer with a crosslinker, so that the average number of bonds to the polyamine monomer is between about 2.05 and about 6; and / or b) producing said polymer by a process wherein the polyamine is initially present in water at a polyamine: water ratio of from about 3: 1 to about 1: 3.

In another aspect, the inventive concept provides a method for preparing an anion-binding polymer that binds a target anion, comprising combining an amine monomer with a crosslinker by a heterogeneous process, wherein the phosphate-binding polymer is characterized in that, at least two of the following particularities: a) a dilation rate of less than about 5; b) less than about 20% by weight of the polymer accessible to non-interacting solutes of molecular weight greater than about twice the MW of the target anion, wherein said percentage is measured in a physiological medium, and c) an interference in binding to the target anion ion of less than about 60% when measured in a gastrointestinal simulator, relative to a non-interfering buffer. In some embodiments the amine monomer is a polyallylamine. In some embodiments the crosslinker is epichlorohydrin.

In another aspect the inventive concept provides an anion-binding polymer that binds a target ion, wherein the polymer is produced by a process comprising crosslinking a polyallylamine by a heterogeneous process, and wherein said polymer is characterized in that at least , two of the following particularities: a) a dilation rate of less than about 5; b) less than about 20% by weight of the polymer accessible to non-interacting solutes of molecular weight greater than about twice the MW of the target anion, wherein said percentage is measured in a physiological medium, and c) an interference in binding to the target anion ion of less than about 60% when measured in a gastrointestinal simulator, relative to a non-interfering buffer. In one embodiment, the polyallylamine is crosslinked by epichlorohydrin.

In another aspect, the inventive concept provides a method for removing an anion from an animal by administering an effective amount of a polymer of the inventive concept to the animal. In some embodiments, the polymer is an anion-binding polymer in which the polymer binds a target anion (eg, phosphate or oxalate), and wherein the polymer is characterized by at least two of the following particularities: a) a dilation rate of less than about 5; b) a pore volume distribution of the gel measured in a physiological medium characterized by a fraction of said pore volume, accessible to non-interacting solutes of molecular weight greater than about twice the MW of the target anion, less than about 20% by weight gel; and c) an ion-binding interference to the target anion of less than about 60% when measured in a gastrointestinal mock relative to a non-interfering buffer. In some embodiments, the target anion of the polymer is phosphate; in some embodiments the phosphate is removed from the gastrointestinal tract; in some embodiments the method of administration is oral. In some embodiments, the animal undergoes at least one disease selected from the group consisting of hyperphosphataemia, hypocalcemia, hyperthyroidism, depressed renal calcitriol synthesis, tetany due to hypocalcemia, renal failure, ectopic soft tissue calcification, and ESRD. In some embodiments, the animal is a human being.

In some embodiments, the polymer is co-administered with at least one of the proton pump inhibitor, calcimimetic, vitamin and its analogues or phosphate binder, e.g. a phosphate binder which is aluminum carbonate, calcium carbonate, calcium acetate, lanthanum carbonate, or SEVELAMER hydrochloride. 16

INCORPORATION BY REFERENCE

All publications and patent applications mentioned in this specification are hereby incorporated by reference to the same extent as if each individual patent application or publication were specifically and individually indicated to be incorporated by reference.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a graph illustrating determination of binding interference by comparing a binding ion of a target ion by a polymer in a non-interfering buffer with binding of a target ion by the polymer in an interfering medium (eg, mock gastrointestinal or fluid collected ex vivo). Figure 2 is a graph illustrating the non-accessible volume of the gel against the radius of the solute probe. Figure 3 is a graph illustrating determination of binding interference for a phosphate-binding polymer (EC172A). Figure 4 is a graph illustrating determination of binding interference for a phosphate-binding polymer (RENAGEL). Figure 5 is a graph of the non-accessible volume versus probe molecular weight for non-interacting probes illustrating the difference between a phosphate-binding polymer of the invention (EC 172A) and a commercially available phosphate binder (RENAGEL ). Figure 6 is a plot of volume not accessible as a function of probe radius for non-interacting probes, illustrating the difference between a phosphate-binding polymer of the invention (EC 172A) and a commercially available phosphate binder (RENAGEL). Figure 7 is a graph illustrating the change in binding capacity by modification of FR-005-144 with chloropropylamine hydrochloride.

The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the particularities and advantages of the inventive concept will be obtained by reference to the following detailed description which defines illustrative embodiments in which the principles of the inventive concept are used, and the appended figures of which: DETAILED DESCRIPTION OF THE INVENTIVE CONCEPT I. Introduction

One aspect of the inventive concept provides polymeric anion-binding materials having one or more of the features of low expansion, high in vivo ion coupling, low interference of interfering ions and / or specific porosity. Another aspect of the inventive concept provides pharmaceutical compositions of the anion-binding polymers, wherein the pharmaceutical composition is a chewable tablet or a liquid formulation. A further aspect of the inventive concept provides methods for preparing or enhancing anion-binding polymers to have one or more of the features of low expansion, high in vivo ion coupling, low interference of interfering ions and / or specific porosity. Yet another aspect of the inventive concept relates to methods of using the anion-binding polymers of the inventive concept for treating conditions in which there is an excess of an ion. In a preferred embodiment, the anion-binding polymers are used to remove target anions from the GI tract. Examples of target anions that can be removed from the GI tract include, but are not limited to, phosphate and oxalate. In another preferred embodiment, the compositions described herein are used in the treatment of hyperphosphataemia, hypocalcemia, hyperparathyroidism, depressed renal calcitriol synthesis, tetany due to hypocalcemia, renal insufficiency, ectopic soft tissue calcification, chronic renal failure and anabolic metabolism. II. Polymers

The inventive concept polymers are characterized by their ability to bind ions. Preferably, the polymers of the inventive concept bind anions, more preferably they bind phosphate and / or oxalate and most preferably they bind phosphate ions. For illustration purposes, anion-binding polymers and, in particular, phosphate-binding polymers will be described; however, it is understood that this disclosure also applies, with appropriate modifications that will be apparent to those skilled in the art, to all ions and 19 solutes. As used herein, a " polymer " an ion, e.g. an anion, or is an " ion-bonding polymer " (eg, a " phosphate-bonding polymer ") when it associates with the ion, generally, though not necessarily, non-covalently, with sufficient strength of association so that at least a fraction of the ions remains bonded under in vitro or in vivo conditions in which the polymer is used for a period of time sufficient to effect removal of the ion from the solution or the organism. A " target ion " is an ion to which the polymer binds, and generally refers to the major ion bound by the polymer or the ion whose binding to the polymer is believed to produce the therapeutic effect of the polymer. A polymer may have more than one target ion. " Link " of an anion, refers to a bond greater than the minimum, i. at least about 0.01 mmol of anion / g of polymer, more preferably at least about 0.05 mmol of anion / g of polymer, yet more preferably at least about 0.05 mmol / , about 0.1 mmol of anion / g of polymer and most preferably at least about 0.5 mmol of anion / g of polymer. The inventive concept provides polymers which are characterized by their selective anion binding; and. In some embodiments, the inventive inventive polymers bind bile acids having a binding activity of less than about 2 mmol / g, preferably less than about 1 mmol / g, more preferably more preferably less than about 0.5 mmol / g, even more preferably less than about 0.3 mmol / g and most preferably less than about 0.1 mmol / g. In some embodiments, the inventive inventive polymers link citrate with a binding activity of less than about 2 mmol / g, preferably less than about 1 mmol / g, more preferably less than about of 0.5 mmol / g, more preferably less than about 0.3 mmol / g, most preferably less than about 0.1 mmol / g. A. Features

Polymers of the inventive concept are characterized by one or more of the following features: 1) low dilation rate; 2) interference at low binding under physiological conditions; 3) a suitable porosity to bind the target anion and to exclude the interfering solutes; 4) an in vivo binding capacity of the target anion sufficient to be effective in therapeutic uses. In some embodiments, the polymer is an anion-binding polymer (eg, a phosphate- and / or oxalate-binding polymer), wherein the polymer is characterized by at least two of the following particularities: 1) a rate of expansion less than about 5; 2) a pore volume distribution of the gel, measured in a physiological medium, characterized in that it is less than about 20% of said pore volume accessible to water soluble non-interacting solutes of molecular weight greater than about twice the PM of the target anion; and 3) an ion-binding interference to the target anion of said polymer of less than about 60% when measured in a gastrointestinal mock relative to a non-interfering buffer. In some embodiments, the polymer is a phosphate-binding polymer characterized by at least one of the following particularities: 1) a dilation rate of less than about 5, preferably less than about 2.5 ; 2) a pore volume distribution of the gel measured in a physiological medium characterized by having less than about 20% of said pore volume accessible to non-interacting solutes of molecular weight greater than about 200; and 3) an interference at ion-to-phosphate binding of less than about 60% when measured in a gastrointestinal mock relative to a non-interfering buffer. In some embodiments, the phosphate-binding polymer has a dilation rate of less than about 2.8, or less than about 2.7, or less than about 2.6. A " physiological medium " is a medium which is isotonic and at neutral pH. In some embodiments the inventive concept provides a phosphate-binding polymer characterized by a dilation rate, measured in isotonic medium at neutral pH, of less than about 5, preferably less than about 2.5, optionally with a mean in vivo phosphate binding capacity of greater than about 0.5 mol / g. In some embodiments, the phosphate-binding polymer has a dilation rate of less than about 2.8, or less than about 2.7, or less than about 2.6. In some embodiments, the inventive inventive polymers bind bile acids having a binding activity of less than about 2 mmol / g, preferably less than about 1 mmol / g, more preferably less than about 0.5 mmol / g, even more preferably less than about 0.3 mmol / g, most preferably less than about 0.1 mmol / g. In some embodiments, the inventive inventive polymers link citrate with a binding activity of less than about 2 mmol / g, preferably less than about 1 mmol / g, more preferably less than about of 0.5 mmol / g, even more preferably less than about 0.3 mmol / g and most preferably less than about 0.1 mmol / g. Preferably, the polymers are comprised of amine monomers.

In general, these particularities are achieved by manipulating one or more factors in the production of the polymer. 22 1) Rate of dilation. The polymers of the inventive concept are crosslinked materials, which means that they do not dissolve in solvents and may at most suffer from dilation in solvents. The rate of dilation in isotonic physiological buffer, representative of the medium of use, i. i.e., the gastrointestinal tract, is typically in the range of about 1.2 to about 100, preferably about 2 to 20. In some embodiments, the inventive concept polymers have a rate of dilation of less than 5, or less than about 4, or less than about 3, or less than about 2.8, or less than about 2.7, or less than about 2, 6, or less than about 2.5. As used herein, " dilation rate " refers to the number of gram of solvent captured by one gram of dry cross-linked polymer when equilibrated in an aqueous medium. When more than one dilatation measurement is performed for a given polymer the mean of the measurements is taken as corresponding to the rate of expansion.

Dilatation rates are measured using a variety of methods: most preferred is the gravimetric method, in which the dry polymer is weighed and added to an excess of liquid. In some cases the liquid may be distilled water; the liquid is preferably an aqueous solution which is isotonic with the plasma; most preferably the liquid is an aqueous solution which is isotonic with the plasma and is buffered at a neutral pH. For example, 0.9% NaCl solution may be used. Phosphate buffered saline (PBS) can also be used. The most preferred physiological medium

for dilation measurements is 0.9% NaCl buffered with 30 mM MES 23 at a pH between about 6.5 and 7.5. The dried polymer (e.g., a phosphate-binding polymer) is generally used in the fully counter-protonated form, e.g. chloride. The polymer is rinsed in the liquid to equilibrium. The rinsed gel is then centrifuged, the decanted supernatant and the heavy wet gel. Care should be taken not to centrifuge at too high a number of g to prevent collapse of the gel. The dilation rate (SR) is calculated as the weight of wet gel minus the weight of dry polymer divided by the weight of the dry polymer.

Another method is the dye method in which an aqueous solution of a very high molecular weight dye is prepared and known to interact with the gel to which an aliquot of dry polymer is added. The weight ratio of the solution to the polymer is adjusted to be close but slightly higher than the expected rate of expansion. As the dye is of very high molecular weight (eg, greater than 200,000 g / mol) it does not cross the gel while the water does so, leading to an increase in the resulting dye concentration, from which the rate of dilatation. An example of a useful dye is fluorescein isothiocyanate modified dextran (FITC). The rate of expansion of a polymer depends on a number of variables such as temperature, ionic strength, polymer charge density, polymer-solvent Flory-Huggins coefficient and crosslink density. Since the ion-binding polymers of the inventive concept are extremely charged polymers (e.g., the phosphate ion-binding polyamines are protonated at the intestinal pH), their dilatation behavior is typically that of polyelectrolyte gels. Although the rate of expansion and the pore size are somewhat related, i. A large swelling rate is generally accompanied by large pores, there is no theoretical basis for accurately estimating the exclusion limit of polyelectrolyte gels. 2) Interference in connection. In some embodiments, the inventive inventive polymers have a binding interference of less than about 70%, more preferably less than about 60%, still more preferably less than about 50%, more preferably less than about 40%, still more preferably less than about 30%, and most preferably less than about 20%, as measured in a gastrointestinal (GI) mock. The phosphate-binding polymers of the inventive concept exhibit binding interference of less than about 70%, more preferably less than about 60%, still more preferably less than about 50%, of a more preferred mode of less than about 40%, still more preferably less than about 30%, and most preferably less than about 20%, as measured in a GI simulator. The " degree of interference in the bond " or " interference in connection, " as used herein refers to the fractional decrease in the binding capacity for the target ion expressed as a percentage observed between a binding experiment in a non-interfering buffer and a gastrointestinal (GI) mock at the same concentration of target anion at equilibrium . An " non-interfering cap, " as used herein refers to a buffer which does not contain one or more solutes that interfere with binding of the target ion and which is buffered to the same pH as the GI mock. A non-interfering buffer is not necessarily free of all interfering solutes, for example, a non-interfering buffer may contain one or both of the ubiquitous gastrointestinal ions, chloride and bicarbonate; if present, these may be at their physiological concentration. An example of a non-interfering buffer is given in Example 1. A " GI simulator " refers herein to a preparation which is designed to mimic the medium of a part of the GI tract upon ingestion of a meal, preferably that part of the GI tract in which the polymer will bind most of the target ion. The GI mock is typically prepared by the method illustrated in Example 1. The target ion should be present in the GI mock at the same concentration (s) as the one (s) used in the non-interfering buffer studies. The degree of interference is easily illustrated by representing the two corresponding binding isotherms, i. i.e. in a GI simulator and in a non-interfering buffer, as shown in Figure 1. An example of determining the interference on the binding using a GI simulacrum is given in Example 1.

It is also possible to measure binding interference by comparing binding of the target ion in digestive fluid collected from individuals, preferably human subjects, with binding of the target ion in non-interfering buffer. If this measurement is carried out, the collected liquids shall be obtained from a number of individuals and the average interference taken as interference in the connection.

It has been found that by carefully choosing the rate of dilation and / or by adjusting the molecular weight cut-off of the gel, the binding capacity in a competitive manner (ie, in vivo or in a GI mock) can be greatly increased relative to other gels with the same polymer composition but whose porosity was not optimized. 26

Importantly, polymers with higher cross-linking and / or crosslinking had less dilation than those with less cross-linking and / or interconnection, and also had binding activity for the target ion (eg, phosphate) which was equal to or greater than the polymers with less cross-linking and / or interconnection. Without wishing to be bound by theory, it is hypothesized that the polymers of the inventive concept exert a sieving effect and fix only solutes of a specific size in solution and exclude other larger species which would otherwise compete with the binding sites within the polymer. The higher molecular weight species include, but are not limited to, inorganic and organic anions, oligopeptides, carbohydrates, bilirubin, lipid mycelia, and lipid vesicles. 3) Porosity. It has been found that it is possible to manipulate the production process of a polymer so that the polymer optimally presents a suitable porosity to bind the target ion (e.g., anions) to which it is intended and to exclude interfering substances. The pore size distribution of polymer gels is obtained by various methods, such as mercury porosimetry, nitrogen adsorption, differential scanning calorimetry or permeation-solute techniques. The latter technique, permeation-solute technique, is most preferred since it tests the gel in a fully hydrated state identical to that prevailing in the medium of use. The solute permeation technique is an indirect method introduced by Kuga (Kuga S.J., J. of Chromatography, 1986,206: 449-461) and consists of measuring the gel partition of solutes of known molecular weights. This method consists of three main steps (Kremer et al., Macromolecules, 1994, 27, 2965-73): 1. Solutions containing dissolved solutes of known molecular sizes and concentrations are contacted with the dilated gel. The molecular sizes of the solutes must cover a considerable range. 2. Dissolve solutes in the gel. The partition of a particular solute depends on the size of the solute and the size distribution of the gel pores. 3. The gel is separated from its surrounding solution and subsequent measurements are made of the concentration of the solutes in the surrounding solution. The decrease in the concentration of each solute relative to its initial concentration is used to calculate the gel pore size distribution.

To separate the molecular exclusion effects from molecular attraction / repulsion effects, the solutes are selected from polymers or oligomers having few or no interactions with the gel type polymer; neutral hydrophilic polymers with a narrow molecular weight distribution, such as polyethylene glycol, polyethylene oxide or dextran, are most suitable. Thus, unless otherwise noted herein, the volumes for exclusion of particular sizes of solutes (also referred to herein as " critical permeation volume ") refer to volumes measured using solutes having essentially no interaction with the polymer for which measurements are taken. 28

Following the experimental protocol and data treatment described in Kremer et al., Macromolecules, 1994, 27, 2965-73, the pore size distribution can be represented as shown in Figure 2. In Figure 2, the Y-axis represents the volume of the dilated gel not accessible to a solute of a certain molecular size. In the example shown in the Figure, small molecules having a size of less than 5 angstroms can penetrate throughout the gel. At the other extreme, polymers with a hydrodynamic radius greater than 1000 angstroms are totally excluded from the gel. In this case the volume not accessible and the volume of gel in equilibrium are equal. The size and molecular weight of the polymers are related through the Mark-Houvink equations which are tabulated for the polymeric solutes used as molecular probes. For example:

Radius (angstrom) = 0.217M ° '498 Dextran

Radius (angstrom) = 0.271 °. 517 Polyethylene glycol

Radius (angstrom) = 0.166M ° 573 Poly (ethylene oxide)

Low molecular weight probes may also be used:

Ureia: Molecular radius 2.5 angstrom Ethylene glycol: Molecular radius 2.8 angstrom Glycerol: Molecular radius 3,1 angstrom Glucose: Molecular radius 4.4 angstrom Sucrose: Molecular radius 5,3 angstrom

Thus, the size of the solute can be converted to molecular weight and vice versa. The size of the solute does not equal the size of the pores; otherwise this would mean that all the liquid in the pores larger than the molecular size of the solute would be available as an accessible volume: this is incorrect because of the effect of the excluded volume, also known as the wall effect.

A direct way to characterize the molecular exclusion limit is to: (i) quantify the partition of molecular probes, (ii) calculate the accessible volume as described above and (iii) normalize it to the volume (or weight) total gel. The desired molecular exclusion limit is achieved by manipulating production variables such as the interconnection of the polymer branches and the crosslinker concentration (see below). Generally, polymers are produced to have a molecular exclusion limit which is based on the ion (eg, anion) to be found and on the probable identity of the interfering substances to be excluded, as well as on the tolerable amount of the inventive concept in the dilation to the intended use for the polymer. In some embodiments of the inventive concept, the ion-binding polymer has a pore volume distribution of the gel (critical permeation volume), defined according to the protocol described above and measured in a physiological medium, of less than about 60% less than about 40%, or less than about 20% of the pore volume of the polymer accessible to non-interacting solutes of molecular weight greater than about twice the MW of the target anion. In some embodiments of the inventive concept, the ion-binding polymer has a gel pore volume distribution (critical permeation volume) of less than about 60%, less than about 40%, or less than about 20% of the pore volume of the polymer accessible to non-interacting solutes of molecular weight greater than about 1.8 times the MW of the target ion. In some embodiments of the inventive concept, the ion-binding polymer has a gel pore volume distribution (critical permeation volume) of less than about 60%, less than about 40%, or less than about of 20% of the pore volume of the polymer accessible to non-interacting solutes of molecular weight greater than about 1.6 times the MW of the target ion. In some embodiments of the inventive concept, the ion-binding polymer has a gel pore volume distribution (critical permeation volume) of less than about 60%, less than about 40%, or less than about of 20% of the pore volume of the polymer accessible to non-interacting solutes of molecular weight greater than about 1.4 times the MW of the target ion. In some embodiments of the inventive concept, the ion-binding polymer has a gel pore volume distribution (critical permeation volume) of less than about 60%, less than about 40%, or less than about of 20% of the pore volume of the polymer accessible to non-interacting solutes of molecular weight greater than about 1.2 times the MW of the target ion. In embodiments, the inventive concept provides a phosphate-binding polymer having a pore volume distribution of the gel (critical permeation volume) defined according to the protocol described above and measured in a physiological medium, less than about 60% , less than about 40%, or less than about 20% of the pore volume of the polymer accessible to non-interacting solutes having a molecular weight greater than about 200, more preferably greater than about 180 , more preferably greater than about 31, about 160, more preferably greater than about 140, and most preferably greater than about 120. 4) Binding ability. The polymers described herein exhibit ion-binding properties, generally anion-binding properties. In preferred embodiments, the polymers exhibit phosphate binding properties. The ion-binding capacity (e.g., phosphate) is a measure of the amount of a particular ion that an ion binder can attach to a given solution. For example the binding capabilities of ion-binding polymers can be measured in vitro and. in water or in physiological saline, or in vivo, e.g. from urinary excretion of ions (e.g., phosphate), or ex vivo, using e.g. liquid collected by aspiration, e.g. chyme obtained from laboratory animals, patients or volunteers. Measurements may be made in a solution containing only the target ion, or at least no other competing solute that will compete with the target ions for attachment to the polymer. In these cases a non-interfering buffer will be used. Alternatively, measurements can be made in the presence of other competing solutes, e.g. other ions or metabolites, which compete with the ions bound for resin binding. The ion-binding capacity for a polymer can be calculated as V * (CinciCeq) / P, expressed in mmol / g, where V is the fixed volume of the solution used, in L; Chalk is the initial concentration of the target ion in the solution in mM; Ceq is the concentration of the target ion at equilibrium in the solution in mM, after a weight P in gram of polymer has been added and allowed to equilibrate. 32

In some embodiments the phosphate fixed polymer. For in vivo use, e.g. in the treatment of hyperphosphataemia, it is desirable for the polymer to have a high phosphate-binding capacity. In vitro measurements of binding capacity do not necessarily translate into in vivo binding capabilities. Thus, it is useful to define binding capacity in terms of in vitro and in vivo capacity. The in vitro phosphate binding capacity of the inventive concept polymers in a non-interfering buffer may be greater than about 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 5, 4.0, 5.0, 6.0, 8.0 or 10.0 mmol / g. In some embodiments, the in vitro phosphate binding capacity of the inventive concept polymers for the target ion is greater than about 0.5 mmol / g, preferably greater than about 2.5 mmol / g, still more preferably greater than about 3 mmol / g, even more preferably greater than about 4 mmol / g, and still more preferably greater than about 6 mmol / g. In some embodiments, the phosphate binding capacity may range from about 0.5 mmol / g to about 10 mmol / g, preferably from about 2.5 mmol / g to about 8 mmol / g and a more preferred from about 3 mmol / g to about 6 mmol / g. Various techniques are known in the art for determining the ability to bind to phosphate. The in vitro phosphate binding capacity of the inventive inventive polymers is measured as described in Example 1 for measuring the binding capacity in a non-interfering buffer.

In some embodiments, the mean ex vivo phosphate binding capacity of the inventive concept phosphate-binding polymers measured in digestive liquids collected from 33 human subjects is greater than about 0.2, 0.3, 4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.2, 1.4, 1.6, 1.8, 2.0, 2.5, 3.0, 4.0, 5.0 or 6.0 mmol / g. Liquids harvested ex vivo are obtained as described in Example 1 from normal subjects, and the binding is measured as for a non-interfering buffer. Average values are taken from about 5-15, or about 15-30, or about 30-60 subjects. In some embodiments, measurements are made on 6-12 subjects.

As used herein, " average phosphate binding capacity in vivo " refers to the binding ability of a polymer as measured in normal human subjects unless otherwise specified, and wherein the binding to the polymer phosphate is measured by the decrease in urinary phosphate excretion combined with the measurement of the phosphate excreted in the feces in the form of free phosphate and bound to the polymer (see below). The mean values are taken from about 5-15, or about 15-30, or about 30-60 subjects. In some embodiments, measurements are made on 6-12 subjects. In some embodiments, the average in vivo phosphate binding capacity of the inventive inventive polymers, preferably measured in human subjects, is at least about 0.3 mmol / g, at least about 0, 5 mmol / g, at least about 0.8 mmol / g, at least about 1.0 mmol / g, at least about 1.5 mmol / g, at least about 2.0 mmol / g, at least about 3.0 mmol / g, at least about 4.0 mmol / g, at least about 5.0 mmol / g, or at least about 6.0 mmol / g. The in vivo binding capacity of the polymer may preferably be determined by measuring the balance of the target ion (eg, phosphate ion) in mammals, preferably in human beings: subjects are given a meal with a controlled content of phosphate and binding polymer, and phosphate and phosphate excreted in feces and urine are monitored. The study comprises a period of elimination followed by a period in which subjects take a daily dose, preferably t.i.d., of phosphate binder, followed by several days without treatment to observe the return to baseline. Phosphate depletion in urine generally coincides with increased phosphate in feces. The moles of phosphate excreted in the faeces subtracted from the base value, and divided by the weight of the binder administered provides a measure of in vivo binding capacity. Unless otherwise noted, measurements " in vivo " mentioned herein use the above protocol. Another method is to measure phosphate binding in vivo and in situ following the protocol set forth in Example 1, wherein the mammals are tubed with a double tube to recover chyme at a particular site of the small intestine. A meal is provided with a certain phosphate content together with a known phosphate binder content and a marker. The label may be a nonabsorbable dye or polymer (e.g., polyethylene glycol), which is then dosed into the chyme to determine the dilution that occurs during the digestion process. The effective concentration of binder is then calculated from the initial concentration at the meal and the dilution rate measured from the marker experiment. The total phosphate in the chyme sample is analyzed. The " soluble " centrifuging the sample and decanting the supernatant and analyzing for the phosphate. The " phosphate " is obtained by the difference between total and soluble phosphates. Two series of experiments are performed on a group of subjects (6-12) taking alternately a placebo (microcrystalline cellulose) or the drug: Binding capacity is obtained by measuring the increase in " bound phosphate " between the two sets of experiments, i.e. with and without drug administration, and dividing by the concentration of binder. The calculation can be done on an individual or group basis. B. Preparation of Polymers

The inventive inventive polymers are prepared by methods known to those skilled in the art; and. ion-binding monomers or their precursors may be copolymerized in the presence of a crosslinking agent; a pre-prepared ion-binding polymer is subsequently cross-linked by a chemical or irradiation reaction; or a precursor polymer is first cross-linked and then reacted to produce the ion-functional functional groups on the polymer.

The polymers are obtained by direct or reverse suspension techniques, emulsion, precipitation, aerosol polymerization or by using crude polymerization / cross-linking methods and size reduction processes, such as extrusion and milling. The processes can be performed as batch, semi-continuous and continuous processes. The rate of expansion, binding interference, binding capacity and PM exclusion limit are affected by at least the following variables of the composition and process: 1 - Concentration of chemical cross-links of the polymer chains. The ratio of (monomer + crosslinker) to the solvent in the crosslinking reaction. The net charge of polymer (at the physiological pH and tonicity of the medium in which it will be used). 4-0 hydrophilic / hydrophobic balance of the polymer backbone. The presence or absence of a coating structure, wherein the coating component restricts the extent of expansion of the inner material.

In the following description the preferred operating ranges for the composition and process variables with crosslinked polyamine materials with phosphate binding properties are exemplified. It will be understood that these are only illustrative conditions, and that the methods described herein may be used in the selection and production of polymers which bind a large range of solutes, as will be apparent to one skilled in the art. 1) Concentration of chemical crosslinks of the polymer chains. The concentration of chemical crosslinks is an important feature that controls the pore dilation and pore distribution properties of the polymer. A convenient way to describe the polymers of the inventive concept is to define an amine-like structural unit and its average number of bonds to the remainder of the polymer. " A " is defined as the amine-type structural unit and " NC " is the average number of bonds from A; NC can be 2, 3, 4 and higher. To form an insoluble gel, NC should generally be greater than 2.

The NC values can then be translated into stoichiometric amine: crosslinker ratios by the following equations:

For low molecular weight monomers, e.g. g., Ν, Ν, Ν ', N'-tetrakis (3-aminopropyl) -1,4-diaminobutane or 1,3-diaminopropane, NC = B-Fb / A, where B is the number of moles of crosslinking agent , Fb is the number of groups in B which react with A to establish a covalent bond, and A is the number of moles of amine.

When the amine type material is of high molecular weight and results from the polymerization of an amine monomer, such as vinylamine, polyethyleneimine, polyvinylamine or polyallylamine, the expression is modified to take account of the two bonds linking the monomeric structural units in the structure center of the polymer. NC is then: NC = (2 ° A + B ° Fb) / A.

Conversely, the molar ratio of crosslinker to the amine can be calculated from the desired NC value by manipulating the above equations:

Low molecular weight amine: B / A = NC'Fb

The following table shows some examples of conversion between NC and the effective ratio of crosslinker to amine, wherein the amine is a high molecular weight material or a small molecule, and wherein the crosslinking material is di- or tri-functional. 38

Amine material Amine type Crosslinkers Fb desired NC Molar ratio A / B Equation Polyalylamine PM epichloro-2 22 0.10 b high hydrin Polyvinylamine PM 1,3- 2 25 0.25 b high dichloropropane Polyethyleneimine PM N-tris (2- 3 22 0.07 b elevated chloroethyl) amine 1,3-Amine 1,3-2,2 1.00 to diaminopropane of low dichloropropane PM Ν, Ν, Ν ', N'-Amine epichloro-2 4,00 to tetrakis (3-hydroxymethylaminopropyl) -1,4-diaminobutane Ν, Ν, Ν ', N'-Amine N-tris (2- -1,4- low ethyl) amine diaminobutane (a): B / A = NC / Fb (b): B / A = (NC-2) / Fb

It was surprisingly found that the selectivity of binding, which reflected in vivo efficacy, was optimized for the NC: in the low range of NC values, the material tended to dilate considerably and exhibited ancillary interference in binding to a stimulant GI. However in the high range, the material had a considerably low intrinsic binding capacity which, obviously, lowered overall performance in vivo. The optimum NC values were found to be between 2.05 and 5 depending on the amine / crosslinker systems.

However, the optimum range to give the desired combination of characteristics in the final polymer depends on the specific monomer and crosslinker used, as well as on other conditions used in the production process, such as the initial concentration of the monomer in the reaction medium, and is a routine experiences.

In some embodiments, the ratio of crosslinker to the total amine groups of the monomers in the polymer is greater than 50 mole%, 60 mole%, 70 mole%, 80 mole% or 90 mole%.

In some embodiments of the inventive concept providing a phosphate-binding polymer containing one or more low molecular weight amine monomers and one or more crosslinkers, NC is greater than about 2, or greater than about 3, or greater than about 4. In some embodiments, the polymers are prepared from crosslinked monomers of N, N, Ν ', Ν' -tetrakis (3-aminopropyl) -1,4-diaminobutane (low PM monomers) with epichlorohydrin (Fb = 2), wherein B / A is from about 2.0 (mol / mol) to about 3.0 (mol / mol) (ie, NC is from about 4 to about 6), or from about 2.3 (mol / mol) to about 2.7 (mol / mol) (ie, NC is from about 4.6 to about 5.4), or about 2, 5 (mol / mol) (i.e., NC is about 5.0). In some embodiments, the polymers are prepared from epichlorohydrin crosslinked N, N, Ν ', Ν'-tetrakis (3-aminopropyl) -1,4-diaminobutane monomers, wherein the initial ratio of the monomer in from about 3: 1 w / w to 1: 3 w / w, or from about 1.5: 1 to about 2: 1 w / w, or about 1: 1, or about 3: 1, wherein B / A is from about 2.0 (mol / mol) to about 3.0 (mol / mol) (eg, NC is from about 4 to about 6), or from about 2.3 (mol / mol) to about 2.7 (mol / mol) (e.g., NC is from about 4.6 to about 5.4), or about 2.5 (mol / mol / mol) (ie, NC is about 5.0). 2) The ratio of (monomer + crosslinker) to the solvent in the crosslinking reaction. High proportions of (monomer + crosslinker) relative to the solvent favor densely crosslinked materials when all other conditions are maintained constant. When a high molecular weight amine is used and when the chain length and polymer concentration are sufficiently large, inter-chain linkages are produced which give rise to many cross-linking nodes as soon as the structure is chemically cross-linked. More generally, for both high and low molecular weight amines, a high proportion of (monomer + crosslinker) relative to the solvent tends to minimize the extent of the side reactions leading to gel defects (eg, intracellular crosslinking yielding cyclic structures, incomplete cross-linking reaction leading to pendent ends).

This condition is determined mainly by the concentrations of both the monomer (e.g., amine) and the crosslinker in the reaction medium. In some embodiments of the inventive concept, the concentration of monomer and crosslinker in the reaction medium is greater than about 20% by weight, preferably greater than 40% by weight, more preferably greater than 60% by weight % by weight. In some embodiments, a (monomer + crosslinker): solvent (e.g., water) ratio of from about 3: 1 to about 1: 3 (w / w) is used. In some embodiments, a (monomer + crosslinker): solvent (e.g., water) ratio of between about 3: 1 to about 1: 1 (w / w) is used. In some embodiments, a (monomer + crosslinker): solvent (eg, water) ratio of about 3: 1, 41 or about 2.5: 1, or about 2.0: 1, or about of 1.5: 1, or about 1: 1 (w / w). The crosslinker can be added at various heights, depending on the polymerization process. In some embodiments, the initial monomer: solvent ratio (prior to addition of crosslinker) is between about 4: 1 to about 1: 1, or between about 3: 1 to about 1: 1; the crosslinker is then added to comprise from about 100 mol% to about 400 mol% of the initial monomer content, or from about 200 mol% to about 300 mol% of the initial monomer content. In some embodiments, the monomer is N, N, Ν ', Ν'-tetrakis (3-aminopropyl) -1,4-diaminobutane and the crosslinker is epichlorohydrin, and the initial monomer: water ratio is between about 4 : 1 to 1: 1, or from about 3: 1 to about 1: 1, or about 1.7, or about 1.73; and the crosslinker is added between about 200 mole% to about 300 mole% of the monomer content, or about 230 mole% to about 270 mole%, or about 250 mole%.

In some embodiments, e.g. the embodiments in which the monomer is a polyallylamine, the amount of monomer is much greater than the amount of the crosslinker (eg, ten times the crosslinker on a molar basis and still more on a weight basis), and the proportions may be expressed as monomer: solvent ratios, bypassing the crosslinker. In some embodiments, the monomer (e.g., polyallylamine) is present in a monomer: solvent ratio of about 3: 1 to about 1: 3. In some embodiments, the monomer is polyallylamine and the crosslinker is epichlorohydrin, wherein the polyallylamine is present in a monomer: water ratio of from about 3: 1 to about 1: 3, and the epichlorohydrin is added to the mixture to about 10 mol% of the total polyallylamine content. 42

Where possible solvent-free processes are even more preferred: in one embodiment the amine and the crosslinker are rapidly mixed and subsequently purely dispersed in a continuous phase, e.g. e.g., water. The crosslinking reaction occurs within the dispersed droplets and is recovered as beads. 3) The net charge of the polymer (at physiological pH and tonicity). The net charge of the polymer is given by the mole content of the ion bond, its intrinsic charge and degree of ionization at physiological pH. The charge density is preferably in the range of 3 to 20 mmol / g, preferably 6 to 15 mmol / g. 4) The hydrophilic / hydrophobic balance of the polymer backbone. The hydrophilic / hydrophobic balance of the polymer allows one to independently control the chemical crosslinking density and the rate of expansion. The rate of expansion is very sensitive to the polymer-solvent interaction parameter% ij as described in the Flory-Huggins formalism (Flory PJ, " Principles of Polymer Chemistry, Cornell, Ithaca Pub., 1953). Increasing the values of% ij to 0.4 and more creates poor solvent conditions for the polymer, which then attempts to minimize the monomer-solvent (water) interaction and consequently dilates much less. This can be achieved by incorporating hydrophobic units in the gel, such as long chain hydrophobic (poly) aromatic substituents, or fluorinated groups. When this strategy is chosen to control the extent of expansion and, consequently, the exclusion limit of the gels, the level of hydrophobic monomers and crosslinkers is from about 0.5 mol% to about 50 mol%, preferably between about 20% and 50%.

In preferred methods, absolute hydrophobicity is quantified by the absolute difference in the log P of the monomers. Quantitatively, the hydrophobic / hydrophilic nature of the monomers can be determined according to the log P of the particular monomers, which is sometimes referred to as the octanol-water partition coefficient. Log P values are well known and are determined according to a standard assay which determines the monomer concentration in a separate water / l-octanol mixture. In particular, it is commercially available, as well as on the Internet, computer programs that allow the estimation of log P values for particular monomers. Some of the log P values in this application were estimated at the http://esc.syrres.com/interkow/kowdemo.htm network site, which provides an estimated log P value for molecules by simply inserting the CAS registration number or a chemical notation. The hydrophobic monomers will typically have a log P value greater than zero and the hydrophilic monomers will typically have a log P value close to or less than zero. In general, the log P of the hydrophobic monomers for purposes of this inventive concept should be at least about 0.5, more preferably at least about 0.75, yet more preferred , at least about 1.0, even more preferably at least about 1.5, and most preferably at least about 2.5. The presence of a coating structure of the core, wherein the liner component restricts the extent of expansion of the core material. Gel particles with coating morphologies are useful in the context of the inventive concept: the coating material may limit the dilation, thereby limiting the exclusion limit, by imposing a chemical resistance to the expansion pressure from the material inside, which could otherwise dilate much more. The coating material may have the same core composition, but with a much higher crosslink density. The design of such enveloping materials and the method for preparing them can be found in U.S. Patent Application No. 10 / 814,789. The coating material may be chemically or physically anchored to the core material. In the first case, the coating can be produced on the core component by chemical means, e.g. by: chemical grafting of the coating polymer into the core using live polymerization from active sites anchored in the core polymer; interfacial reaction, i.e. and, a localized chemical reaction on the surface of the core particle, such as interfacial polycondensation; and using block copolymers as suspending agents during the synthesis of core particles. Interfacial reaction and the use of block polymers are preferred techniques when using chemical methods. Typically, in the interfacial reaction path, the periphery of the core particle is chemically modified by reacting small molecules or macromolecules at the core interface. For example, an amine-containing ion-binding nuclear particle is reacted with a polymer containing amine-reactive groups, such as epoxy groups, isocyanate, activated esters, halides to form a cross-linked coating around the core. 45

In another embodiment, the coating is first prepared using interfacial polycondensation or solvent coacervation to produce capsules. The inside of the capsule is then filled with core forming precursors to construct the core within the capsule film.

In some embodiments using the block copolymer approach, an amphiphilic block copolymer may be used as a suspending agent to form the core particle in a reverse or direct suspension particle formation process. When a water-in-oil reverse suspension process is used, the block copolymer then comprises a first block soluble in the continuous oil phase and another hydrophilic block contains functional groups which can react with the core polymer. When added to the aqueous phase, together with the core forming precursor and the oil phase, the block copolymer is located at the water-in-oil interface and acts as a suspending agent. The hydrophilic block reacts with the core material or co-reacts with the core forming precursors. After the particles have been isolated from the oil phase, the block copolymers form a thin film covalently attached to the surface of the core. The chemical nature and length of the blocks may be modified to alter the permeation characteristics of the film relative to the solutes of interest.

When the coating material is physically adsorbed to the core material, well-known microencapsulation techniques such as solvent coacervation, fluidized bed spray coating, or multiemulsion processes may be employed. A preferred microencapsulation method 46 is the fluidized bed spray coating in the Wurster configuration. In yet another embodiment, the coating material acts only temporarily to retard dilation of the core particle while in the mouth and esophagus, and optionally disintegrates in the stomach or duodenum. In this case the film is selected to hinder the transfer of water to the core particle, creating a layer of high hydrophobicity and very low permeability to water.

Thus, in one aspect, the inventive concept provides a method for selecting an ion-binding polymer, wherein the polymer contains a monomer and a crosslinker, and wherein the polymer has at least one of the particularities of a) a rate of dilation less than about 5; b) a pore volume distribution of the gel measured in a physiological medium characterized by a fraction of said pore volume, accessible to non-interacting solutes of molecular weight greater than about twice the MW of the target anion, less than about 20% by weight gel; and c) an ion-binding interference to the target anion of less than about 60% when measured in a gastrointestinal mock, relative to a non-interfering buffer: i) by modifying the following composition and process variables 1) the ratio of crosslinker to monomer ; 2) the ratio of (monomer + crosslinker) to the solvent in the reaction medium; 3) the net charge of the polymer at physiological pH and tonicity; and / or (4) the hydrophilic / hydrophobic balance of the polymer backbone (ii) evaluating the dilation, porosity and interference properties of the resulting polymer ion; and iii) selecting a polymer having at least one of the foregoing particularities.

In another aspect the concept of the invention provides a method for improving the therapeutic and / or aptitude for administration and / or pharmaceutical properties of a polyamine polymer comprising at least one of the following steps: a) crosslinking said polymer with a crosslinking agent such that the average number of bonds to the polyamine monomer is between about 2.05 and about 6; and / or b) producing said polymer by a process wherein the polyamine is initially present in water at a polyamine: water ratio of from about 3: 1 to about 1: 3. B. Monomers

Any suitable monomers and crosslinkers may be used in the polymers of the inventive concept. When the polymer alloys phosphate or oxalate, the polymer generally comprises a polyamine and a crosslinker. The polyamines include functional amine monomers such as those described in U.S. Patents No. 5496545; 5,676,775; 6509013; 6132706; and 5,968,499; and in U.S. Patent Applications 10/806495 and 10/701385. These patents and patent applications 48 are hereby incorporated by reference in their entirety.

In some embodiments, the inventive concept provides ion-binding polymers containing crosslinked amine units. In some of these embodiments, the polymers are characterized by one or more of the features of low expansion, high in vivo ion coupling, low interference of interfering ions and / or specific porosity. Polymers, including homopolymers and copolymers, with crosslinked amine basic structural units are referred to herein as cross-linked amine polymers. The basic amine structural units in the polymer may be separated by the same length or different lengths of (or intercalated) binding moieties. In some embodiments, the polymers comprise the basic structural units of an amine and an intercalated linker. In other embodiments, the multiple amine units are separated by one or more binding units.

A useful monomer in the polymers of the inventive concept comprises an amine of formula I

(1) wherein each n is independently equal to or greater than 3; m is equal to or greater than 1; and each R 1 is independently H or optionally substituted alkyl or aryl or is attached to a vicinal R 1 to form an optionally substituted alicyclic, aromatic or heterocyclic group. In one embodiment, the inventive concept is a cross-linked amine polymer comprising an amine of Formula I as described, wherein the amine is cross-linked with a crosslinking agent.

Preferred amines of formula I include:

n: 3.4, or 5

In one aspect, the inventive concept provides methods of treating an animal, including a human, using the inventive concept polymers. One embodiment of this aspect is a method for removing phosphate from the gastrointestinal tract of an animal by administering an effective amount of a cross-linked amine polymer, wherein said polymer comprises an amine of formula I.

A second monomer useful in the polymers of the inventive concept comprises an amine of formula II

Fig.

N. wherein: p is 1, 2, 3 or 4; each R 1 is independently H or optionally substituted alkyl or aryl or is attached to a vicinal R 1 to form an optionally substituted alicyclic, aromatic or heterocyclic group; R2 and R3 are each independently H, optionally substituted alkyl or aryl, provided that when p = 1, R2 and R3 are not H and when p = 2, 3 or 4, R2 and R3 are H, alkyl or -C (R 1) 2 -R 4 -N (R 2) 2, wherein R 4 is a bond or methylene; In addition, in some embodiments, the amines of formula II include amines wherein p is greater than 4. In various embodiments, p may be greater than 8, greater than 12, greater than 16, or higher than 20. In other embodiments, p may be less than 25, less than 20, less than 15 or less than 10. In one embodiment the inventive concept is a cross-linked amine polymer comprising an amine of formula II, as described, wherein the amine is crosslinked with a crosslinking agent.

Preferred amines of formula II include:

nh2 V nh2 51

One embodiment of the inventive concept is a method for removing phosphate from the gastrointestinal tract of an animal by administering an effective amount of a crosslinked amine polymer, wherein said polymer comprises an amine of formula II.

A third useful monomer in the inventive concept polymers comprises an amine of formula III

(III) wherein q is 0, 1 or 2; and each R 1 is independently H or optionally substituted alkyl or aryl or is attached to a vicinal R 1 to form an optionally substituted alicyclic, aromatic or heterocyclic group. In one embodiment the inventive concept is a cross-linked amine polymer comprising an amine of formula III, as described, wherein the amine is crosslinked with a crosslinking agent.

Preferred amines of formula III include:

One embodiment of the inventive concept is an animal method for removing phosphate from the gastrointestinal tract of a patient by administering an effective amount of a cross-linked amine polymer, wherein said polymer comprises an amine of formula III.

A fourth monomer useful in the polymers of the inventive concept comprises an amine of formula IV

(IV) wherein each n is independently equal to or greater than 3; each r is, independently, 0, 1 or 2; and each R 1 is independently H or optionally substituted alkyl or aryl or is attached to a vicinal R 1 to form an optionally substituted alicyclic, aromatic or heterocyclic group. In one embodiment the inventive concept is a cross-linked amine polymer comprising an amine of formula IV as described, wherein the amine is crosslinked with a crosslinking agent.

A preferred amine of formula IV includes:

53

One embodiment of the inventive concept is a method for removing phosphate from the gastrointestinal tract of an animal by administering an effective amount of a crosslinked amine polymer, wherein said polymer comprises an amine of formula IV.

A fifth monomer useful in the inventive concept polymers comprises an amine of formula V

wherein each n is independently equal to or greater than 3; each r is, independently, 0, 1 or 2; and each R 1 is independently H or optionally substituted alkyl or aryl or is attached to a vicinal R 1 to form an optionally substituted alicyclic, aromatic or heterocyclic group. In one embodiment the inventive concept is a cross-linked amine polymer comprising an amine of formula V as described, wherein the amine is crosslinked with a crosslinking agent.

Preferred amines of formula V, as used in accordance with the present invention, are: η 2ν η 2η

νη2 η: 3, 4 or 5 νη2

One embodiment of the inventive concept is a method for removing phosphate from the gastrointestinal tract of an animal by administering an effective amount of a cross-linked amine polymer, wherein said polymer comprises an amine of formula V.

A sixth useful monomer in the inventive concept polymers comprises an amine of formula VI

(VI) wherein each m is independently equal to or greater than 3. In one embodiment the inventive concept is a crosslinked amine polymer comprising an amine of formula VI as described, wherein the amine is crosslinked with an agent of crosslinking.

One embodiment of the inventive concept is a method for removing phosphate from the gastrointestinal tract of an animal by administering an effective amount of a cross-linked amine polymer, wherein said polymer comprises an amine of formula VI.

The amines represented by the general formulas I-VI may be synthesized by methods well known in the art. These synthetic techniques include the catalytic conversion of alcohols, reductive amination of carbonyl compounds, Michael additions and hydrogenation of nitriles (see, eg, Karsten Eller et al., Ullmann's Encyclopedia of Industrial Chemistry 2002 by Wiley-VCH Verlag GbH &; Co. KGaA). Also commercially available are various amine and / or amine monomers and intercalated linkers.

In one embodiment, an amine useful in the present inventive concept, tetramethylenetetraamine, shown below, is synthesized by catalytic hydrogenation of the commercially available diaminomaleonitrile (DAMN):

Catalyst H2 .... r.-ΠΠ.ΤΓ n - nh2 nh2 nh2 nh2

The amines which may be used in the present inventive concept are not limited, but are typically small amines which serve as monomers or parts of monomeric units for the polymerization reactions. In some embodiments, the monomers are low molecular weight monomers, i. monomers having a molecular weight of less than 200 g / mol.

In embodiments of the inventive concept, the monomers are non-polymeric, e.g. non-polymeric amines. As used herein, " polymer " encompasses a molecule of high relative molecular mass, the structure of which essentially comprises the multiple repetition of units derived, either orally or conceptually, of molecules of low relative molecular mass. 56 of the

Examples of amines which are suitable for the polymer synthesis of the present inventive concept include, but are not limited to, the amines shown in Table 1. TABLE 1

B-SM-23-DA Diamina h2n / x /% nh2 61.1 B-SM-22-DA Diamine h2n / x /% nh2 61.1 B-SM-20-TeA Tetramine ' DA Diamina / NH, h2n 2 88.15 B-SM-24-DA Diamine r- {H} NH2 74,13 B-SM-25-DA Diamine XX HjN NHZ 88.15 B-SM- '-1' -NHj 129.21 B-SM-27-DA Diamine 114.19 B-SM-28-TA Triamine NHa Cr ** "Xfz * 2Ha NH, 196,08 B-SM-29-TA Triamine «Λγ'γΝΗ, NyM NH, 125.13 B-SM-31-DA Diamine Gr * N- ma 184.07 57

Label Type Structure PM (g / mol) B-SM-32-DA Diamina A MHj NH2 136.2

Additional amine monomers that may be used in the inventive inventive polymers include vicinal amine units. The polymer may be a homopolymer including vicinal amine basic structural units or is a copolymer comprising one or more vicinal amine basic structural units and other monomers such as acrylates, methacrylates, acrylamides, methacrylamides, vinyl esters, vinyl amides, olefins, styrenes, etc. The size of the polymer may vary between, e.g. about 500 to about 1000,000 Daltons.

A vicinal amine monomer useful in the inventive concept polymers is the monomer shown in formula VII:

R

(I.e.

Wherein n is zero, one or more than 1, each R is independently a suitable chemical group which complements the nitrogen valence, and each R 'is independently H, alkyl or amino.

In another embodiment, the polymer is characterized by a respective structural unit having the formula

R

or a copolymer thereof, wherein n is zero, one or more than 1, each R is independently a suitable chemical group which complements the valency of the nitrogen, each R 'is independently H, alkyl or amino, and X' is a counterion organic or inorganic charge.

Preferred polymers of formula VIII include:

59

The polymers of the present inventive concept also include polymers characterized by a respective unit having the formula?

R

Formula IX wherein n is zero, one or more than 1, each R is independently a suitable chemical group which complements the nitrogen valence, each R 'is independently H, alkyl or amino, and X- is an organic or inorganic counterion negatively charged.

In one embodiment, the R groups of vicinal nitrogen atoms are connected to each other to give a structure as depicted in formula X

Wherein X is a bond, alkyl, alkylamino, alkylcarbonyl, alkenyl, aryl or heterocyclyl.

In the polymers described herein, n is zero, one or more than 1. In preferred embodiments, n is 0-5, still more preferably n is zero or 1. The value of n 'depends on the properties desired for the polymer, the potential use of the polymer and the synthetic techniques used. The pendant nitrogen atom in the formulas VII, VIII, IX and X may be attached to atoms such as C, H, O, S, P and N such that pendant groups are nitrous, nitro, nitroxide radical, nitrone, nitrene , isocyanate, carbazide, hydrazino, diazo, imine, amidine, guanidine, sulfamate, phosphoramidate and heterocycle groups.

Examples of suitable R groups include H, halogen, R ", CO₂H, CO₂R ", C (= NR "), CN, CONH2, CONR ", OR ", S03R ",Si; 3 and P (O) (OR ") 2. The R " Suitable alkyl groups include H, and optionally substituted alkyl, acyl, alkylamino, alkenyl, heterocyclyl and aryl groups. Preferred R 'is H, methyl or amino.

Substituents for the R " may be ionic entities with oxygen, nitrogen, phosphorus or sulfur. Examples of substituents are the carboxylate, sulfonate, sulfamate, sulfone, phosphonate, phosphazene, phosphoramidate, quaternary ammonium groups, or amine groups, e.g. e.g., primary and secondary alkyl or aryl amines. Examples of other suitable substituents include hydroxyl, alkoxy, carboxamide, sulfonamide, halogen, alkyl, aryl, hydrazine, guanadine, urea, and carboxylic acid esters.

Preferred R groups include H and the following groups -CH3

t • 0-ch2chs -ch2ch2nh2 (ch2ch2nh2 * ^

CH

NH-NH 2

Vo ♦ " Ντ

T

T (Alkyl or Aryl) (Alkyl or Aryl) ΌΗ

h2n nh

R (Alkyl or Aryl) N

H2N NH

Oh

T (Alkyl or Aryl)

T

HO

ÒH HO OH

(Alkyl or Aryl) I-OH

the point of attachment for groups R

The negatively charged counterions, X-, may be organic ions, inorganic ions or a combination thereof. Inorganic ions suitable for use in this inventive concept include halides (especially chloride), carbonate, bicarbonate, sulfate, bisulfate, hydroxide, nitrate, persulfate and sulfite. Suitable organic ions include acetate, ascorbate, benzoate, citrate, dihydrogencitrate, hydrogencitrate, oxalate, succinate, tartrate, taurocholate, glycocholate and cholate. Preferred is chloride or carbonate.

In a preferred embodiment, the counterion has no detrimental side effect to the patient and is selected so as to have a therapeutic or nutritional benefit to the patient.

Another monomer for use in the polymers of the inventive concept is of the formula XI shown below,

Rw

nr2 where R " is H or CH 3, and R has the same meaning as above. Preferred structures of formula XI are those wherein R = H.

In one embodiment, the polymer is a copolymer wherein one of the basic building blocks is a monomer as described herein.

The copolymers of the present inventive concept may be alternating or random copolymers. In general, monomers that can be copolymerized with the amine precursors include one or more monomers selected from the group consisting of styrene, substituted styrene, alkyl acrylate, substituted alkyl acrylate, alkyl methacrylate, alkyl methacrylate acrylonitrile, methacrylonitrile, acrylamide, methacrylamide, N-alkylacrylamide, dialkylacrylamide, N-alkylmethacrylamide, N, N-dialkylmethacrylamide, N, N-isoprene, butadiene, ethylene, vinyl acetate, N-vinylamide, maleic acid derivatives, vinyl ether, allyl, methallyl and combinations thereof. Functionalized versions of these monomers may also be used. Specific monomers or comonomers which may be used in this inventive concept include, but are not limited to, methyl methacrylate, ethyl methacrylate, propyl methacrylate (all isomers), butyl methacrylate (all isomers), 2- ethylhexyl, isobornyl methacrylate, methacrylic acid, benzyl methacrylate, phenyl methacrylate, methacrylonitrile, (X-methylstyrene, methyl acrylate, ethyl acrylate, propyl acrylate (all isomers), butyl acrylate (all isomers ), 2-ethylhexyl acrylate, isobornyl acrylate, acrylic acid, benzyl acrylate, phenyl acrylate, acrylonitrile, styrene, glycidyl methacrylate, 2-hydroxyethyl methacrylate, hydroxypropyl methacrylate (all isomers), methacrylate of hydroxybutyl acrylate (all isomers), Ν, Ν-dimethylaminoethyl methacrylate, Ν, Ν-diethylaminoethyl methacrylate, triethylene glycol methacrylate, itaconic anhydride, itaconic acid, glycidyl acrylate (all isomers), hydroxybutyl acrylate (all isomers), Ν, Ν-dimethylaminoethyl acrylate, Ν-diethylaminoethyl acrylate, triethyleneglycol acrylate, methacrylamide, N- N-butyl methacrylamide, N-methylolmethacrylamide, N-ethylolmethacrylamide, N-tert-butylacrylamide, N-butylacrylamide, N-methylolacrylamide, N-ethylolacrylamide, 4-acryloylmorpholine, vinylbenzoic acid, N-tert-butylmethacrylamide, (all isomers), diethylaminostyrene (all isomers), 64α-methylvinylbenzoic acid (all isomers), diethylamino-oc-methylstyrene (all isomers), p-vinylbenzenesulfonic acid, p-vinylbenzenesulfonic acid sodium salt, methacrylate trimethoxysilylpropyl methacrylate, methacrylate methacrylate methacrylate methacrylate methacrylate methacrylate, diethoxysilylpropyl methacrylate, tribethoxysilylpropyl methacrylate, dimethoxy metilsililpropilo, dietoximetilsililpropilo, dibutoximetilsililpropilo, diisopropoximetilsililpropilo, dimetoxissililpropilo, dibutoxissililpropilo methacrylate methacrylate, diisopropoxissililpropilo methacrylate, trimetoxissililpropilo acrylate, trietoxissililpropilo acrylate, tributoxissililpropilo acrylate, dimetoximetilsililpropilo acrylate, dietoximetilsililpropilo acrylate, dibutoximetilsililpropilo acrylate, diisopropoximetilsililpropilo of acrylate, diisopropoxysilylpropyl acrylate, maleic anhydride, N-phenylmaleimide, N-butylmaleimide, N-vinylformamide, N-vinylacetamide, allylamine, methallylamine, allyl alcohol, methyl vinyl ether, ethyl vinyl ether, butyl vinyl ether, butadiene, isoprene, chloroprene, ethylene, vinyl acetate and combinations thereof. Preferred monomers or comonomers are acrylamide, dimethylacrylamide, N-vinylformamide, N-vinylacetamide, vinyl acetate, methyl acrylate and butyl acrylate.

Other monomers that can be used in the polymer of the inventive concept include: X.

N /

R \

Wherein each R is independently H or a substituted or unsubstituted alkyl, such as a lower alkyl group (eg, having 1 to 5 carbon atoms inclusive), alkylamino (eg, having 1 to 5 ring atoms carbon, inclusive, such as ethylamino) or aryl (eg, phenyl); R X-

wherein each R is independently H or a substituted or unsubstituted alkyl group (eg, having 1 to 5 carbon atoms inclusive), alkylamino (eg, having from 1 to 5 carbon atoms inclusive, such as ethylamino ) or aryl (eg, phenyl), and each X- is a negatively charged exchangeable cation.

Another suitable monomer is a structure of formula

Wherein R is H or a substituted or unsubstituted alkyl group (eg, having 1 to 5 carbon atoms inclusive), alkylamino (eg, having from 1 to 5 carbon atoms inclusive, such as ethylamino) or aryl (eg, phenyl).

Another suitable monomer is a structure of the formula X-

wherein each R 1 and R 2 are independently H or a substituted or unsubstituted alkyl group (eg, having 1 to 5 carbon atoms inclusive), and alkylamino (eg, having 1 to 5 carbon atoms inclusive, such as ethylamino) or aryl (eg, phenyl), and each X 'is a negatively charged exchangeable counter ion. In one embodiment, at least one of the R groups is a hydrogen atom.

Another suitable monomer is a structure of formula

wherein each R 1 and R 2 is independently H, a substituted or unsubstituted alkyl group containing 1 to 20 carbon atoms, an alkylamino group (eg, having from 1 to 5 carbon atoms inclusive, such as ethylamino), or an aryl group containing 6 to 12 atoms (eg phenyl).

Another suitable monomer is a structure of the formula x-

Ri? Wherein each R 1 and R 2 and R 3 is independently H, a substituted or unsubstituted alkyl group containing 1 to 20 carbon atoms, an alkylamino group (eg, having 1 to 5 carbon atoms, inclusive, such as ethylamino), or an aryl group containing 6 to 12 atoms (eg, phenyl), and each X- is a negatively charged exchangeable counter ion.

In each case of these monomers, the R groups may carry one or more substituents. Suitable substituents include therapeutic anionic groups, e.g. quaternary ammonium groups or amine groups, e.g. primary and secondary alkyl- or aryl amines. Examples of other suitable substituents include hydroxyl, alkoxy, carboxamide, sulfonamide, halogen, alkyl, aryl, hydrazine, guanadine, urea and carboxylic acid esters, for example.

The negatively charged counterions, X-, may be organic ions, inorganic ions or a combination thereof. Inorganic ions suitable for use in this inventive concept include halide (especially chloride), carbonate, bicarbonate, sulfate, bisulfate, hydroxide, nitrate, persulfate and sulfite. Suitable organic ions include acetate, ascorbate, benzoate, citrate, dihydrogencitrate, hydrogencitrate, oxalate, succinate, tartrate, taurocholate, glycocholate and cholate.

Guanidino group containing polymers are also useful as compositions which can be produced by the processes described herein to have the desired properties and which binds anions, such as phosphate and oxalate. Such polymers are described in U.S. Patent Nos. 6,132,706; and 5,968,499, which are hereby incorporated by reference in their entirety. Briefly, guanidino groups are attached to a polymeric structure. The nature of the polymer backbone is not of primary importance since the binding effect is due to the guanidino groups. Preferred polymers in which the cross-linking and other factors can be controlled include polymers having a cross-linked polyethylene backbone with divinylbenzene. Also, polymers having an inorganic backbone, e.g. polyphosphazene polymers. The polymers may be copolymers derived from two or more different types of monomers. Other examples of useful polymers are carbohydrate polymers including cellulose and agarose. The guanidino groups are attached to the central structure of the polymer by chemical bonding through the terminal NH group of the guanidino group (NH 2 -C (= NH) -NH-). The chemical binding of the guanidino groups to the polymer backbone may be either directly or through some form of the group acting as a " spacer " through which it is attached to the central structure of the polymer. Various binding forms may be used, the preferred forms varying with the basic type of polymer. For example, alkylene groups of 1-4 carbon atoms, amide groups, ether groups or a combination thereof may be used. The preferred mode of attachment of the guanidino groups to the central structure of the polymer will obviously depend on the nature of the core structure but for simplicity's sake, direct bonding between atoms of the central structure and the NH group of the guanidino group is preferred where possible.

Methods of preparing guanidino-containing polymers will be apparent to one skilled in the art, but e.g. g., can be prepared following the teachings of Schnaar, RL and Lee, YC, 1975, Biochemistry 14, 1535-1541, incorporated herein by reference in its entirety, which describes a method for binding biologically active ligands to a polymer matrix, or the polymers may also conveniently be prepared by reaction with a polymer containing amino groups attached to the polymer backbone of (a) 3,5-dimethylpyrazole-1-carboxamidine nitrate, (b) S-methylthiouronium sulfate or ) O-methylpseudourea hydrogensulfate.

Preferred monomers of the inventive concept are amines. Highly preferred monomers for use in the inventive inventive polymers include allylamine, vinylamine, ethyleneimine, methylene-1,3-diamino-propane, and Ν, Ν, Ν ', N'-tetrakis (3-aminopropyl) -1,4 diaminobutane, 1,2,3,4-tetraaminobutane, formula 1 and formula 2, wherein formula 1 and formula 2 are the following structures:

70

In some embodiments, the inventive concept polymers are comprised of one or more amine monomers and one or more crosslinkers wherein the polymer is produced by a process in which the amine is present in the solvent prior to crosslinking in an amine: solvent ratio from about 3: 1 to about 1: 3 and the total content of crosslinkers added to the reaction mixture is such that the average number of bonds to the amine monomers is between about 2.05 and about 6, or between about 2.2 and about 4.5. In some embodiments, the inventive inventive polymers are a phosphate-binding polymer comprised of one or more amine monomers and one or more crosslinkers wherein the polymer is produced by a process wherein the total content of crosslinkers added to the reaction mixture is such that the average number of bonds to the amine monomers is between 2.2 and 4.5. In preferred embodiments, the amine monomer is selected from the group consisting of 1,3-diaminopropane, and N, N, Ν ', Ν'-tetrakis (3-aminopropyl) -1,4-diaminobutane, and wherein the The crosslinking agent is selected from the group consisting of 1,3-dichloropropane and epichlorohydrin. In some embodiments, the inventive inventive polymers are comprised of one or more amine monomers and one or more crosslinkers, wherein the amine monomers are not polyallylamine monomers and / or the crosslinkers are not epichlorohydrin.

In some embodiments, e.g. in phosphate-binding polymers, it is desirable to maintain the ratio of chloride to the amine of the final polymer below certain levels. In some embodiments, this is about 0 to about 35 mol%, preferably about 0 to about 15 mol%. The monomers can be selected according to this criterion. D. Crosslinkers The crosslinker includes those described in U.S. Patents No. 5496545; 5,676,775; 6509013; 6132706; and 5,968,499; and U.S. Patent Applications 10/806495 and 10/701385.

Crosslinking agents are typically compounds having at least two functional groups which are selected from a halogen group, carbonyl group, epoxy group, ester group, acid anhydride group, acid halide group, isocyanate group, vinyl group and chloroformate group . The crosslinking agent may be attached to the carbon backbone or pendant nitrogen of the amine polymer. Examples of crosslinkers that are suitable for the synthesis of the polymers of the present inventive concept include, but are not limited to the crosslinkers shown in Table 2. TABLE 2

Label Structure PM X-EP-1 ΪΧ / CI 92.52 X-EP-2 174.19 X-EP-3 tn ** 0 72 (continued)

Label Structure PM X-EP-4 302.37 X-EP-5 297.27 X-EP-6 O0 277.32 X-EP-7 86.09 X-EP-80 202.25 X-Cl-1 1 H-NMR 184.41 X-Cl-2 175.06 X-Cl-3 112.99 X-Cl-4 H2 Cl / * X / N > / xC1 Cf 240.99 X-Cl-6 127.01 73 (continued)

Label Structure PM X-AC-1 ° 203.02 X-AC-2 "A0 203.02 X-AC-3 V 265.48 α ct X-AC-4 154.98 X-AH-1 Ák 198, 13 X-AH-2 OMe X-AH-3 Â ± 112.08 X-Mc-1 H-8 H 0 168.2 X-Mc-2 11 or 118.16 X-Mc-3 | / 249.27 74 (cont'd)

75 (continued) Label Structure PM X-ME-7? 194.19 X-ME-8 O 178.14 X-ME-9 / O 108.53

Examples of suitable crosslinking agents are diacrylates and dimethacrylates (eg ethylene glycol diacrylate, propylene glycol diacrylate, butylene glycol diacrylate, ethylene glycol dimethacrylate, propylene glycol dimethacrylate, butylene glycol dimethacrylate, polyethylene glycol dimethacrylate, polyethylene glycol diacrylate), methylene bisacrylamide, methylene bisethacrylamide, ethylene bisacrylamide, ethylene bismethacrylamide, ethylidene bisacrylamide, divinylbenzene, bisphenol A dimethacrylate, bisphenol A diacrylate, diepoxides, dihalides, diisocyanates, diacyl chlorides, dianhydrides and dimethyl esters. preferred include diglycidyl, 1,3-dichloropropane, 1,2-dibromoethane, toluene diisocyanate, ethylene bisacrylamide

Examples of cross-linking agents epichlorohydrin, 1,4-butanediol ether 1,2-ethanediol diglycidyl ether, 1,2-dichloroethane, 1,3-dibromopropane, succinyl dichloride, dimethylsuccinate, acryloyl chloride, methyl acrylate, and dianhydride pyromellitic. E. Polymerization Polymerization can be accomplished by methods known in the art, examples of which are illustrated in detail in the Examples described herein. As described above, the polymerization conditions can be engineered to produce polymers having the desired characteristics. The crosslinking reaction is performed either in crude solution (i.e., using the pure amine and pure crosslinker) or in dispersed medium. The crosslinking reaction leading to the formation of the gel can be carried out using a variety of processes; These processes fall into two categories: i) homogeneous processes in which the functional amine precursor (small molecule of amine or polyamine of high molecular weight) is soluble in the continuous phase, and wherein the gel obtained by the cross-linking reaction is recovered as a crude gel or gel paste in said continuous phase. The crude gel process describes the situation where the entire solvent is trapped in the gel network forming a mass which is then transformed into smaller particles using extrusion, grinding and related methods. When a crude process is used, the solvents are selected so that they co-dissolve the reactants and do not interfere with the cross-linking reaction of the amine. Suitable solvents include water, low boiling alcohols (methanol, ethanol, butanol), dimethylformamide, dimethylsulfoxide, acetone, methyl ethyl ketone and the like. A gel paste is typically obtained in which the viscosity of the reaction medium is in the low range and a high shear rate whereby pieces of gel are produced which remain in suspension in a paste form. Ii) heterogeneous processes in which the functional amine precursor (small molecule of amine or polyamine of high molecular weight) is rendered insoluble in the continuous phase so as to form drops or dispersed particles, which then undergo a cross-linking reaction, forming a pearl or irregularly shaped particles held in suspension in said continuous phase.

Homogeneous processes may be impractical for crosslinked materials having restricted expansion rates, such as those contemplated in this inventive concept: the typical crosslinking level for the desired range of expansion rate and pore size distribution generally induces a very short gelation time and high local viscosity, both of which are impractical in industrial scale manufacture.

A preferred mode of synthesis for the present inventive concept is the use of heterogeneous processes. Such processes are also referred to as polymerization in dispersed media and include reverse suspension, direct suspension, precipitation polymerization, emulsion polymerization and microemulsion polymerization, reaction in aerosols and the like. The continuous phase may be selected from apool solvents such as toluene, benzene, hydrocarbon, halogenated solvents, supercritical carbon dioxide and the like. With a direct suspension or emulsion process water may be used, although the saturated salt solutions are also useful for separating the amine and crosslinker into a separate drop phase, as described in U.S. Patent No. 5414068. The monomer precursors can be dispersed neat or as a solution in the continuous phase. The amine and the crosslinker are preferably introduced in two separate steps, wherein the amine is first dispersed as droplets and the crosslinker is subsequently added to the reaction medium and migrates to the dispersed phase. The crosslinking reaction occurs within the droplet phase without causing any significant increase in the viscosity of the dispersion. This has the advantage of dissipating the heat generated by the exothermic reaction while ensuring good homogeneity of the gel within the beads. A preferred synthetic mode comprises the steps of: i) solubilizing the amine monomer or amine polymer in water ii) neutralizing a portion of the amine with an acid, such as HCl, iii) dispersing said amine solution in a solvent immiscible with water to form an emulsion iv) adding the crosslinker to the emulsion in a stepwise addition v) allowing the crosslinking reaction to proceed to completion vi) removing the water by distillation vii) isolating the beads by filtration viii) washing and drying

In this process the polymer particles are obtained as spherical beads, the diameter of which is preferably controlled in the range of 5 to 500 micrometers, preferably 25 to 250 micrometers. In some of these embodiments the beads have an average diameter of less than 40 micrometers.

Thus, in one aspect, the inventive concept provides a method for preparing an anion-binding polymer that binds a target anion, comprising combining an amine monomer 79 with a cross-linker in a heterogeneous process, wherein the phosphate-binding polymer is characterized by at least two of the following particularities: a) a dilation rate of less than about 5, or less than about 4.5, or less than about 4, or less than about 3 ; b) less than about 20% by weight of the polymer accessible to non-interacting solutes of molecular weight greater than about twice the MW of the target anion, wherein said percentage is measured in a physiological medium, and c) an interference in binding to the target anion ion of less than about 60% when measured in a gastrointestinal simulator, relative to a non-interfering buffer. In some embodiments the amine monomer is a polyallylamine. In some embodiments the crosslinker is epichlorohydrin.

In another aspect the inventive concept provides an anion-binding polymer that binds a target ion, wherein the polymer is produced by a process comprising crosslinking a polyallylamine by a heterogeneous process, and wherein said polymer is characterized in that at least , two of the following particularities: a) a dilation rate of less than about 5, or less than about 4.5, or less than about 4, or less than about 3; b) less than about 20% by weight of the polymer accessible to non-interacting solutes of molecular weight greater than about twice the MW of the target anion, wherein said percentage is measured in a physiological medium, and c) an interference in binding to the target anion ion of less than about 60% when measured in a gastrointestinal simulator, relative to a non-interfering buffer. In one embodiment, the polyallylamine is crosslinked by epichlorohydrin. 80

As discussed above, the molar ratios of crosslinker to the amine control the extent of gel material formed as well as its crosslink density. Too low a proportion can lead to incomplete cross-linking and formation of soluble oligomers, whereas too low a proportion can produce an extremely tight network with few binding properties. The amine component may be a single or a combination of various amines, and the same applies to the crosslinking component. An optimization for any new combination of amines and crosslinkers may be required, since the functionality of any of them can influence the extent of gel formation and the dilatation characteristics. In some embodiments, e.g. in the low molecular weight monomers crosslinked crosslinkers having a Fb of 2, the molar ratios of crosslinker to the amine (B / A) are in the range of about 0.2 to about 10, of a preferably about 0.5 to about 5, and most preferably about 0.5 to about 2. These ratios may be adjusted by treating a high molecular weight or low molecular weight amine monomer, and / or based on the Fb number of the crosslinker (see discussion and table above).

In some cases the polymers are cross-linked after polymerization. One method for achieving such crosslinking involves reacting the polymer with bifunctional crosslinkers, such as epichlorohydrin, succinyl dichloride, diglycidyl ether of bisphenol A, pyromellitic dianhydride, toluene diisocyanate and ethylenediamine. A typical example is the reaction of poly (ethyleneimine) with epichlorohydrin. In this example the epichlorohydrin (1 to 100 parts) is added to a solution containing polyethyleneimine (100 parts) and heated to promote the reaction. A typical example is the reaction of vicinal polyamine with epichlorohydrin. In this example the epichlorohydrin (1 to 200 parts) is added to a solution containing vicinal polyamine (100 parts) and heated to promote the reaction. Other methods of inducing cross-linking in already prepolymerized materials include, but are not limited to, exposure to ionizing radiation, ultraviolet radiation, electron beams, radicals and pyrolysis. The crosslinking reaction is carried out in batches or semi-continuously. In the latter mode, the amine or the crosslinker is added as the starting charge and the co-reactant is then metered for a certain period of time. In one embodiment, a soluble prepolymer is first prepared by adding the entire amine monomer component and then continuously adding a fraction of the crosslinker, which gives a syrup. The syrup is then emulsified into droplets in a continuous oil phase and the remaining crosslinking moiety is added to form crosslinked beads. Where the crosslinker is an alkyl halide compound, a base may be used to retain the acid formed during the reaction. Inorganic or organic bases are suitable. NaOH is preferred. The ratio of base to crosslinker is preferably from about 0.5 to about 2.

In some embodiments the polymers are post-aminated (post-reaction with 3-chloropropylamine). In this embodiment, a first reaction is carried out between an amine monomer and a crosslinker to form a gel, the gel is then reacted with an aminoalkyl halide in which the amine-alkyl groups are chemically bound to the gel through the replacement of the halide by functional amine gels.

All of the polymers described herein may further be crosslinked and impregnated with anion, e.g. phosphate. In one embodiment the target anion (e.g., phosphate or oxalate) is present during the polymerization and is then washed out once the crosslinking reaction is complete. The method is referred to as &quot; printing &quot; and tends to increase the chemical affinity of the gel for the anionic solute by creating &quot; molded &quot; in the gel having high binding recognition for a certain anion. Examples of gels with phosphate pockets are described, e.g. in Fujiwara et al, Analytical Sciences April 2000, vol, 16, 407, and in ACS symposium series 703, &quot; Molecular and Ionic Recognition with Imprinted Polymers, Bartsch RA and Maeda M. Editors, 1998, 315. Typically the anion is present in a molar ratio to the amine (expressed as nitrogen atom) of from about 10% to about 100%, more preferably about 10% to about 60%, in a very about 30% to about 50%. Most preferably, the anion is introduced into the acid form (e.g., phosphoric acid, oxalic acid) and the amine as the free base, so as to form the ammonium / anion salt in situ. The crosslinking is then performed as described above using the appropriate ratio of crosslinker to the amine to provide the desired characteristics for the gel in terms of rate of expansion, critical permeation volume and interference on binding. The gel obtained immediately after cross-linking is then washed extensively in extremely acidic (e.g., pH <2) or extremely basic (e.g., pH> 12) medium to remove the impregnated anion, and then washed again with neutral medium.

By keeping all parameters equal (eg, the ratio of amine 83 to crosslinker, ratio of monomer to solvent) the impregnation method described herein generally increases the binding capacity by a factor of 1.1, 1.1 or even 1.5. III. Pharmaceutical Compositions

In one aspect the inventive concept provides pharmaceutical compositions. In one embodiment, the pharmaceutical compositions are chewable tablets. In another embodiment, the pharmaceutical compositions are liquid formulations.

The pharmaceutical compositions of the present inventive concept include compositions in which the inventive concept polymers, e.g. crosslinked amine polymer, are present in an effective amount, e.g. in an amount effective to achieve a therapeutic and / or prophylactic benefit. The effective amount effective for a particular application will depend on the patient (e.g., age, weight), the condition being treated and the route of administration. Determination of an effective amount is within the skill of the person skilled in the art, especially in light of this disclosure. The amount effective to be used in humans can be determined from animal models. For example, a dose may be formulated for humans to achieve circulating and / or gastrointestinal concentrations which have been found to be effective in animals. 84

The pharmaceutical compositions comprise the polymer, e.g. crosslinked amine polymers, one or more pharmaceutically acceptable carriers, diluents or excipients, and optionally further therapeutic agents.

The pharmaceutical compositions to be used according to the present inventive concept may be formulated in a conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate the processing of the active compounds into preparations which may be used pharmaceutically. The appropriate formulation depends on the route of administration chosen. Suitable techniques for preparing pharmaceutical compositions from the amines are well known in the art, e.g. Gennaro AR (ed), Remington's Pharmaceutical Sciences, 20th Edition, Lippincott, Williams and Wilkins, Baltimore MD (2001), which is hereby incorporated by reference in its entirety.

The current pharmaceutical compositions are generally prepared by known procedures using well known and readily available ingredients. In preparing the compositions of the present inventive concept, the ion-binding polymer, e.g. a phosphate-binding polymer, may be present alone, may be combined with a carrier, diluted with a carrier or enclosed within a carrier which may be in the form of a capsule, sachet, paper, or other container. When the carrier serves as a diluent, it may be a solid, semi-solid or liquid material which acts as a vehicle, excipient or medium for the polymer. Thus, the compositions may be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, syrups, aerosols (as a solid or in a liquid medium), hard or soft gelatin capsules, sterile powders packaged and similar. Preferred formulations are chewable tablets and liquid formulations. Examples of carriers, excipients and diluents which may be used in these formulations as well as in others include foods, beverages, lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum arabic, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose , polyvinylpyrrolidone, cellulose, methylcellulose, methyl hydroxybenzoates, propyl hydroxybenzoates, propyl hydroxybenzoates and talc.

In another aspect of the inventive concept, the anion-binding polymer (e.g., phosphate) is formulated as a free, non-counter-aminated amine. Short-term and long-term studies have shown that patients on hemodialysis treated with Renagel (polyallylamine hydrochloride) have significantly lower serum bicarbonate levels than patients treated with calcium-containing phosphate binders (i.e., no chloride). It has been demonstrated (Brezina B. et al., Kidney International, vol 66, suppl. 90 (2004), 39-45) that SEVELAMER hydrochloride (Trade name of the pharmaceutical active ingredient Renagel) induces an acid overload causing acidosis. Acidosis can have serious side effects for this type of patient. In another embodiment, the crosslinked amine polymer is a polyamine polymer wherein the content of the chloride in the polymer is less than about 40 mol% of the amine group content, more preferably less than about 20% molar ratio of the amine group content and still more preferably less than about 5% of the amine group content. Most preferably, the polymer is essentially free of chloride. 86 A. Chewable tablets

In some embodiments the inventive concept polymers are provided as pharmaceutical compositions in the form of chewable tablets. Patient compliance is currently recognized as one of the major limiting factors for patients to adhere to recommendations in the treatment of ionic imbalance disorders, such as hyperphosphataemia. For example in the treatment of hyperphosphataemia using current phosphate-binding polymers, such as RENAGEL, recent surveys have shown that patients have to take on average nine to ten 800 mg tablets per day, taking 25% of the patient population still daily doses higher doses of twelve to fifteen pills. Renagel is in the form of tablets to be ingested and is administered with quantities of fluid necessary to ingest the tablets, which overload ESRD patients who are under fluid restriction. The low adherence of patients due to high daily doses stands out as a factor that clearly affects the acceptance of this class of drugs.

Pharmaceutical formulations which are easier to take would be desirable. Although administration of drugs through a chewable tablet would be extremely advantageous in many cases, use has been limited since formulators have encountered difficulties in achieving satisfactory sensory characteristics. When chewing a tablet the following sensory parameters are important: sand sensation, tooth adhesion, powder sensation, mouth feel and overall palatability. 87

Chain chew tablets are used mainly in areas where significant amounts of active substances have to be administered and include over-the-counter products such as vitamins, antacids, laxatives and pain medications. Prescription chewable products include prenatal vitamins and antibiotic and antiviral chewable products that require oral administration of large doses. While often large, the geometry has to be optimized to facilitate chewing and &quot; hardness &quot; suitable for chewing. Round shapes of bevelled ends with height / diameter ratios of about 0.3 to 0.4 are common.

In addition to the active ingredient the following types of excipients are customarily used: a sweetener to provide the necessary palatability, and a binder when the former is unsuitable to provide a sufficient tablet hardness; a lubricant to minimize friction effects on the mold wall and facilitate ejection of the tablet; and in some formulations a small amount of a disintegrant is added to facilitate chewing. Generally, the excipient levels in the currently available chewable tablets are in the range of 3-5 times the active ingredient (s) while the sweeteners constitute the bulk of the inactive ingredients.

An important consideration in the design of a chewable tablet containing an ion-binding polymer is the rate of dilation of the polymer. Since the inventive concept provides for slow-diluting polymers, they may be used in chewable formulations without the unpleasant and sometimes dangerous side effects that accompany the chewable tablets of the higher dilatation polymers. One example of a raised dilatation material that causes difficulties during oral administration and which potentially results in clogging and blockage of the esophagus is Psyllium. Psyllium comes from the crushed seeds of the Plantago ovata plant, a plant species native to parts of Asia, Mediterranean regions of Europe and North Africa and is generally used as a laxative in the USA. Typically Psyllium dilates 35-50 times and has to be ingested with many fluids. Insufficient fluid intake during administration may lead to fiber to dilate and result in obstruction or even rupture of the esophagus. Psyllium is contraindicated in patients with dysphagia and / or a narrow esophagus. The present inventive concept provides chewable tablets which contain a polymer or polymers of the inventive concept and one or more pharmaceutical excipients suitable for the formulation of a chewable tablet. The polymer used in chewable tablets of the inventive concept preferably has a rate of dilation as they pass through the oral cavity and through the esophagus of less than about 5, preferably less than about 4, more preferably less than about 3, more preferably less than 2.5, and most preferably less than about 2. In some embodiments the polymer is an anion-binding polymer such as a polymer of bonding to phosphate or oxalate; in a preferred embodiment, the polymer is a phosphate-binding polymer. The tablet comprising the polymer, combined with suitable excipients, provides acceptable organoleptic properties such as mouth feel, taste and adhesion to the teeth, and at the same time does not pose any risk of obstruction of the esophagus after chewing and contact with saliva.

In some aspects of the inventive concept, the polymer (s) provides mechanical and thermal properties which are generally performed by excipients, thereby decreasing the amount of such excipients necessary for formulation. In some embodiments the active ingredient (eg, polymer, preferably an anion-binding polymer) constitutes more than about 30%, more preferably more than about 40%, even more so. Preferably more than about 50% and most preferably more than about 60% by weight of the chewable tablet, the remainder of suitable excipient (s) consisting of. In some embodiments the polymer, e.g. anion-binding polymer, comprises about 0.6 g to about 2.0 g of the total weight of the tablet, preferably about 0.8 g to about 1.6 g. In some embodiments, the polymer, e.g. an anion binding polymer, comprises more than about 0.8 g of the tablet, preferably more than about 1.2 g of the tablet and most preferably more than about 1 , 6 g of the tablet. The polymer is produced to have suitable strength / friability and particle size to provide the same qualities for which excipients are often used, e.g. suitable hardness, good mouth feel, compressibility and others. The particle size for the polymers used in the chewable tablets of the inventive concept is less than about 80, 70, 60, 50, 40, 30 or 20 microns in average diameter. In preferred embodiments, the particle size is less than about 80, more preferably less than about 60-90 and most preferably less than about 40 micrometers.

Useful pharmaceutical excipients in the chewable tablets of the inventive concept include a binder, such as microcrystalline cellulose, colloidal silica and combinations thereof (Prosolv 90), carbopol, providone and xanthan gum; a flavoring such as sucrose, mannitol, xylitol, maltodextrin, fructose or sorbitol; a lubricant, such as magnesium stearate, stearic acid, sodium stearyl fumarate and vegetable based fatty acids; and optionally a disintegrant, such as croscarmellose sodium, gellan gum, hydroxypropyl cellulose ether, sodium starch glycollate. Other additives may include plasticizers, pigments, talc and the like. These additives and other suitable ingredients are well known in the art; see, e.g. Gennaro AR (ed), Remington's Pharmaceutical Sciences, 20th Edition.

In some embodiments the inventive concept provides a pharmaceutical composition formulated as a chewable tablet, comprising a phosphate-binding polymer and a suitable excipient. In some embodiments the inventive concept provides a pharmaceutical composition formulated as a chewable tablet, comprising a phosphate-binding polymer, a filler and a lubricant. In some embodiments the inventive concept provides a pharmaceutical composition formulated as a chewable tablet, comprising a phosphate-binding polymer, a filler and a lubricant, wherein the filler is selected from the group consisting of sucrose, mannitol, xylitol, maltodextrin, 91 and fructose and sorbitol, and wherein the lubricant is a magnesium salt of a fatty acid, such as magnesium stearate. The tablet may be of any size and shape compatible with chewing and disintegrating in the mouth, preferably in a cylindrical form, having a diameter of about 10 mm to about 40 mm and a height of about 2 mm to about of 10 mm, most preferably a diameter of about 22 mm and a height of about 6 mm.

In one embodiment the polymer has a transition temperature greater than about 30øC, preferably greater than about 50øC.

In another embodiment, the polymer is pre-formulated with a low molecular weight, high melting point / Tg excipient such as mannitol, sorbose, sucrose to form a solid solution in which the polymer and the excipient are intimately mixed. Mixing methods such as extrusion, spray drying, freeze drying, lyophilization or wet granulation are useful. The indication of the mixing level is given by known physical methods, such as differential scanning calorimetry or dynamic mechanical analysis.

Methods for preparing chewable tablets containing pharmaceutical ingredients, including polymers, are known in the art. See, e.g. European Patent Application No. EP373852A2 and U.S. Patent No. 6475510, and Remington's Pharmaceutical Sciences, which are hereby incorporated by reference in their entirety. 92 B. Net formulations

In some embodiments the inventive concept polymers are provided as pharmaceutical compositions in the form of liquid formulations. In some embodiments the pharmaceutical composition contains an ion-binding polymer dispersed in a suitable liquid excipient. Suitable liquid excipients are known in the art; see, e.g. Remington's Pharmaceutical Sciences. IV. Methods of treatment

In another aspect, the inventive concept provides methods of treating ionic imbalance disorders. The term &quot; ionic imbalance disorders &quot; as used herein refers to conditions in which the level of an ion present in the body is abnormal. In one embodiment, the inventive concept provides methods of treating a phosphate imbalance disorder. The term &quot; phosphate imbalance disorder &quot; as used herein refers to conditions in which the level of phosphorus present in the body is abnormal. An example of a phosphate imbalance disorder includes hyperphosphatemia. The term &quot; hyperphosphatemia &quot; as used herein refers to a condition in which the phosphorus element is present in the body at a high level. Typically, a patient is often diagnosed as suffering from hyperphosphatemia if the level of phosphate in the blood is, e.g. more than about 4.5 milligrams per deciliter of blood and / or the glomerular filtration rate is reduced, e.g. g., by more than about 20%. 93

Thus, for example, the inventive concept provides methods for removing an anion from an animal by administering an effective amount of a polymer of the inventive concept to the animal. In some embodiments, the polymer is an anion-binding polymer wherein the polymer binds a target anion (eg, phosphate or oxalate), and wherein the polymer is characterized by at least two of the following particularities: a) a dilation rate less than about 5; b) a pore volume distribution of the gel measured in a physiological medium characterized by a fraction of said pore volume accessible to non-interacting solutes, of molecular weight greater than about twice the MW of the target anion, less than about 20% by weight gel; and c) an ion-binding interference to the target anion of less than about 60% when measured in a gastrointestinal mock relative to a non-interfering buffer. In some embodiments, the target anion of the polymer is phosphate; in some embodiments the phosphate is removed from the gastrointestinal tract; in some embodiments the method of administration is oral. In some embodiments, the animal undergoes at least one disease selected from the group consisting of hyperphosphataemia, hypocalcemia, hyperthyroidism, depressed renal calcitriol synthesis, tetany due to hypocalcemia, renal failure, ectopic soft tissue calcification, and ESRD. In some embodiments, the animal is a human being. It will be understood that any polymer described herein may be useful for attaching an anion to an animal and / or for treating conditions caused by an ionic imbalance in an animal. In preferred embodiments, the polymer is a phosphate-binding polymer wherein the polymer is characterized by at least one of the following particularities: a) a dilation rate of less than about 5, preferably less than which is about 2.5; B) a pore volume distribution of the gel measured in a physiological medium characterized by a fraction of said pore volume, accessible to non-interacting solutes of molecular weight greater than about 200, less than about 20% by weight of gel; and c) an interference at ion-to-phosphate binding of less than about 60% when measured in a gastrointestinal mock, relative to a non-interfering buffer. In some embodiments, the rate of dilation is less than about 2.8, or less than about 2.7, or less than about 2.6.

Other diseases which may be treated with the methods, compositions and kits of the present inventive concept include hypocalcemia, hyperparathyroidism, hungry bone syndrome, depressed renal synthesis of calcitriol, tetany due to hypocalcemia, renal insufficiency and ectopic calcification in soft tissues including calcifications in joints , and in lung, kidney, connective and myocardial tissues. Likewise, the present inventive concept can be used to treat patients with ESRD and undergoing dialysis, including for the prophylactic treatment of any of the foregoing.

Also, the polymers described herein can be used as an adjuvant of other therapies, e.g. g, those using phosphorus feed intake, dialysis of inorganic salts and / or other polymeric resins.

The compositions of the present inventive concept are also useful in the removal of chloride, bicarbonate, iron ions, oxalate and bile acids from the gastrointestinal tract. The polymers removing oxalate ions find use in the treatment of an oxalate imbalance disorder, such as oxalosis or hyperoxaluria which increases the risk of renal calculi formation. Polymers which remove chloride ions find use, e.g. e.g., in the treatment of acidosis, heartburn, acid reflux disease, acid stomach or gastritis. In some embodiments, the compositions of the present inventive concept are useful for removing fatty acids, bilirubin, and related compounds. Some embodiments may also bind to and remove high molecular weight molecules such as proteins, nucleic acids, vitamins or cellular debris. The present inventive concept provides methods, pharmaceutical compositions and kits for the treatment of animals. The term &quot; animal &quot; or &quot; animal subject &quot; as used herein includes humans as well as other mammals. One embodiment of the inventive concept is a method of removing phosphate from the gastrointestinal tract of an animal by administering an effective amount of at least one of the cross-linked amine polymers described herein. The term &quot; treatment &quot; and their grammatical equivalents as used herein include achieving a therapeutic benefit and / or a prophylactic benefit. By therapeutic benefit is meant the eradication, amelioration or prevention of the underlying disorder to be treated. For example in a patient with hyperphosphatemia, the therapeutic benefit includes the eradication or amelioration of the underlying hyperphosphatemia. Also, a therapeutic benefit is achieved by eradicating, ameliorating or preventing one or more of the physiological symptoms associated with the underlying disorder whereby an improvement in the patient is observed, although the patient may continue to be affected by the underlying disorder. E.g., administration of the cross-linked amine polymers described herein to a patient suffering from renal failure and / or hyperphosphatemia provides a therapeutic benefit not only when there is a decrease in serum phosphate in the patient, but also when an improvement in the patient is observed in relation to other disorders that accompany renal failure and / or hyperphosphatemia such as ectopic calcification and renal osteodystrophy. For prophylactic benefit, for example the cross-linked amine polymers may be administered to a patient at risk of developing hyperphosphatemia or to a patient who describes one or more of the physiological symptoms of hyperphosphatemia, even if a diagnosis of hyperphosphataemia has not yet been made. E.g./r the inventive concept polymers may be administered to a patient with chronic renal disease in whom hyperphosphatemia has not been diagnosed.

The dosages of the polymer, e.g. crosslinked amine polymers in animals will depend on the disease to be treated, the route of administration and the physical characteristics of the animal to be treated. In some embodiments in which crosslinked amine polymers are used, the dosage levels of the crosslinked amine polymers for therapeutic and / or prophylactic uses may be from about 1 g / day to about 30 g / day. It is preferred that these polymers are administered together with the meals. The polymers may be administered once a day, twice a day or three times a day. The preferred dosage range is from about 2 g / day to about 20 g / day and an even more preferred dosage range is from about 3 g / day to about 7 g / day. The dose of the polymers described herein may be less than about 50 g / day, preferably less than about 40 g / day, yet more preferably less than about 30 g / day, of an even more preferred mode of less than about 20 g / day, and most preferably is less than about 10 g / day.

Preferably, the ion-binding polymers, e.g. crosslinked amine polymers used for therapeutic and / or prophylactic benefits may be administered alone or in the form of a pharmaceutical composition as described herein. For example the crosslinked amine polymers of the present inventive concept may be co-administered with other active pharmaceutical agents depending on the pathology to be treated. Examples of pharmaceutical agents that may be co-administered include, but are not limited to, proton pump inhibitors, calcium mimetics (e.g., cinacalcet), Vitamin D and their analogues and phosphate binders. Examples of suitable phosphate binders include, but are not limited to, aluminum carbonate, calcium carbonate, calcium acetate (PhosLo), lanthanum carbonate (Fosrenol) and Renagel. Such co-administration may include simultaneous administration of the two agents in the same dosage form, simultaneous administration into separate dosage forms and separate administration. For example for the treatment of hyperphosphatemia, the cross-linked amine polymers may be co-administered with calcium salts which are used to treat hypocalcemia resulting from hyperphosphatemia. The calcium salt and the polymer may be formulated together in the same dosage form and administered simultaneously. Alternatively, the calcium salt and the polymer may be administered simultaneously, both agents being presented in separate formulations. In another alternative, the calcium salt may be administered soon followed by the polymer, or vice versa. In the separate administration protocol, the polymer and the calcium salt may be administered separated from a few minutes, or separated from a few hours, or separated from several days. The polymer may be administered by injection, topically, orally, transdermally or rectally. Preferably, the polymer or the pharmaceutical composition comprising the polymer is administered orally. The oral form in which the polymer is administered may include powder, tablet, capsule, solution or emulsion. The effective amount may be administered in a single dose or in a series of separate doses for appropriate time intervals, such as hours. The inventive concept also provides methods for removing anionic pollutants from waste water by contacting the waste water with an anion-binding polymer of the inventive concept, wherein the anionic pollutants, e.g. phosphate, are adsorbed to the polymer. V. Kits

In yet another aspect, the present inventive concept provides kits for the treatment of anion imbalance disorders, e.g. for the treatment of phosphate imbalance disorder. These kits comprise a polymer or polymers described herein and instructions explaining the use of the kit according to the various methods and approaches described herein. These kits may also include information, such as references to scientific literature, packaging materials, clinical trial results and / or summaries thereof and others, which indicate or establish the activities and / or advantages of the composition. This information can 99 studies be based on the results of various studies, and. using experimental animals involving in vivo models and studies based on human clinical trials. The kits described herein may be provided, marketed and / or donated for the purpose of promotion to health care providers, including physicians, nurses, pharmacists, form officers, and the like. The kits for cosmetic use can be provided, marketed and / or donated for promotional purposes directly to consumers.

All publications and patent applications mentioned in this specification are hereby incorporated by reference to the same extent as if each individual patent application or publication were specifically and individually indicated to be incorporated by reference.

It will be apparent to one skilled in the art that many changes and modifications may be made to the disclosure herein without departing from the spirit or scope of the appended claims.

EXAMPLES

Example 1: Phosphate binding measurement protocols

In this Example various protocols are described for measuring the ability of a polymer to bind an anion (in this case, phosphate). 100

Measurements of the phosphate binding capacity in a non-interfering buffer

An aliquot of dried polymer of weight P (g) was mixed under gentle shaking with a fixed volume, V (L), of the following buffer: 20 mM H 3 PO 4, 80 mM NaCl, 100 mM MES (morpholinoethanesulfonic acid) sodium salt and a pH of 6.5. When single measurements of binding were made, the last buffer was used. When multiple measurements were taken, e.g. for the representation of a binding isotherm, the concentration of phosphate in the buffer was varied. The initial concentration of phosphate ion is referred to as PiniCiai (mM). The solution may be referred to as a non-interfering buffer since it contains no competing solutes competing with the phosphate ions for attachment to the polymer resin. After equilibration of the resin, the solution was decanted by centrifugation and the supernatant was analyzed for residual phosphate concentration by ion chromatography, Peq (mM). Binding capacity was calculated as V * (Piniciai-Peq) / P, expressed in mmol / g as indicated in the tables for the corresponding polymers.

Ability to attach in a gastrointestinal simulacrum

This process is designed to mimic the conditions of use of a phosphate-binding polymer in a GI tract and to measure the binding characteristics of the polymer to the phosphate (target solute) in the presence of other metabolites (competing solutes). A liquid meal was artificially digested in the presence of pepsin and gastric juice to produce a gastrointestinal (GI) mock. The addition sequence of 101 enzymes and the pH profile were controlled so that the digestion process was simulated to the level of the jejunum:

The following components were added one at a time in the following order: Powdered Milk 291 g, Beneprotein 72.8 g, Dextrose 152 g, Polycosis 156 g, NaCl 17.6 g - 2.5 L of H2 O, to dissolve (they were stirred vigorously but avoided foaming). After the NaCl had dissolved, 240 g of Corn oil was added. The volume was made up to 4 L with H 2 Odd. The mixture was stirred vigorously for 2 hours. At this point the pH was -6.4. Then 153 mL of 3M HCl was added dropwise to a final pH of 2.0 (150 mL). The mixture was stirred for 15 minutes, after which the pH was ~2.1. 800 ml of pepsin was then added in 10 mM HCl to a final concentration of 1 mg / ml. The mixture was stirred at RT for 30 minutes, after which the pH was -2.3. 5 L of a stock solution of Pancreatin and Biliary Salts in 100 mM NaHCO 3, pH 8.4 to a final concentration of 0.3 mg / mL of Pancreatin and 2 mg / mL of Biliary Salts were then added. The mixture was stirred for 120 minutes at ambient temperature, whereupon the pH was -6.5. The mock meal was retained at -80øC for up to one month prior to use.

An aliquot of the GI simulacrum was centrifuged and the supernatant was analyzed for phosphate. The phosphate binding assay was similar to that described above with non-interfering buffer, except using liquid from the GI mock. 102

Ability to connect in an ex-vivo aspirator

Using a tube placed in the lumen of the small intestine, healthy subjects were given a meal of the same composition as prepared for the GI mockery described above and then chyme aliquots were harvested. The subjects were intubated with a double polyvinyl tube with a bag containing a heavy amount of mercury at the end of the tube to facilitate movement of the tube into the small intestine. Using fluoroscopy to guide placement, a concentric double tube aspiration opening was located in the stomach and the other aperture was located in the Treitz Ligament (in the upper jejunum).

After correct tube placement, 550 mL was infused into the liquefied test meal (supplemented with a marker, polyethylene glycol (PEG) - 2 g / 550 mL) in the stomach through the gastric opening at a rate of 22 mL per minute. It took about 25 minutes for the entire meal to reach the stomach, simulating the time required to eat normal meals.

The jejunal chemo was aspirated from the tube whose lumen was placed in the Treitz Ligament. This fluid was collected continuously at intervals of 30 minutes over a period of two and a half hours. This gave 5 samples which were mixed, measured on a volume basis and lyophilized.

A phosphate binding assay was performed on ex-vivo liquids collected by aspiration. The phosphate binding process was similar to that described above with non-interfering buffer, except that the liquid collected ex-vivo (after reconstitution of the freeze-dried material in the appropriate amount of deionized water) was used. The phosphate binding capacities in the ex-vivo harvested liquid were calculated in the same manner as in the GI mock experiments.

Example 2: Crosslinked polymer libraries prepared in a crude solution process and measurement of the phosphate binding capacity

Preparation of Polymer Libraries

The following five examples each cover a library comprising up to 24 crosslinked polymers. The polymers were prepared in discontinuous reactors organized in a 4x6 matrix format. Each reactor had a volume of 350 microliters or 3 ml was magnetically stirred and the temperature controlled. In a typical procedure, the amine, the crosslinkers, solvents and optionally the base were supplied by a robot to each reactor, optionally under stirring. The reactors were then sealed and heated to the indicated temperature for 15 hours. The reactor array was then removed, and crosslinked polymer buffers were transferred into glass vials, ground, repeatedly washed with deionized water, and lyophilized. The five libraries are identified below in Table 3 together with the corresponding reaction conditions used in their preparation. TABLE 3

Example Library identification Reaction temperature (° C) Reactor volume (microliters) 1 100275 85 350 2 100277 60 350 3 100279 80 350 4 100353 80 350 5 100384 80 3000

Measurements of the phosphate binding capacity in a non-interfering buffer

The binding capacities for the phosphate ion were also determined for each of the polymers in the libraries. See Example 1 for the process.

Results

Tables 4-8 indicate the materials and quantities used in the preparation of the polymers from each of the 5 libraries together with the phosphate binding capacities measured in a non-interfering buffer for the prepared polymers. The intakes correspond to the weights of chemicals used in each reaction well in mg along with the phosphate binding capacity of the obtained polymer gel (white indicates that no crosslinked gel was obtained in that particular reaction). 105 TABLE 4

Library: Plate 3 (ID; 100275) Unit: mg

Row Column Water B-SM-22-DA X-Cl-3 NaOH DMSO Phosphate Binding (mmol / g) 1 1 128.51 67.74 51.63 9.14 0.00 1 2 130.70 57.94 61.82 10.94 0.00 1 3 132.33 50.61 69.43 12.29 0.00 1 4 133.59 44.93 75.33 13.33 0.00 3.042 1 5 134.60 40 , 39 80.04 14.17 0.00 0 1 6 135.43 36.69 83.89 14.85 0.00 0 2 1 136.42 32.26 88.50 15.66 0.00 3.703 2 2 137.05 29.41 91.45 16.19 0.00 3,624 2 3 137.58 27.03 93.93 16.63 0.00 2.858 2 4 138.03 25.00 96.03 17.00 0, 00 2,566 2 5 138.42 23.26 97.84 17.32 0.00 2,761 2 6 138.76 21.74 99.42 17.60 0.00 2.82 3 1 132.04 64.98 49, 52 17.53 34.60 3 2 134.77 55.13 58.82 20.82 47.26 3 3 136.79 47.87 65.67 23.25 57.22 3 4 138.34 42.30 70 , 93 25.11 65.27 3.087 3 5 139.57 37.90 75.09 26.58 71.91 2.946 3 6 140.56 34.32 78.47 27.78 77.48 2.535 4 1 141.75 30.06 82.48 29.20 79.73 2.674 4 2 142.50 27.35 85.04 30.11 90.45 3.038 4 3 143.13 25.09 87.18 30.86 97.98 2.895 4 4 143.66 23.17 88.99 31.50 103.56 2.571 4 5 144.12 21.52 90.54 32.05 107.86 2.636 4 6 0.0 0 0.00 0.00 0.00 0.00 5.374 TABLE 5

Table 1 Table 2 Table 1 Table 2 Table 2 Table 2 Table 2 Table 2 Table 2 Table 2 Table 2 Table 2 Table 2 Table 2 Table 2 Table 2 Table 2 Table 2 Table 2 Table 2 Table 2 Table 2 Table 2 Table 1 Column 95 0.00 DMF 1 2 124.02 107.66 16.36 0.00 0.00 1 3 124.33 104.74 19.59 0.00 0.00 1 4 124.63 101.98 22.65 0.00 0.00 1 5 124.91 99.35 25.55 0.00 0.00 4,183 1 6 125.17 96.86 28.31 0.00 0.00 4,237 2 1 125.59 92.98 32.61 0.00 0.00 4,631 2 2 125.89 90.08 35.81 0.00 0.00 4.594 2 3 126.18 87.37 38.81 0.00 0.00 4,667 2 4 126, 45 84.81 41.64 0.00 0.00 4.586 2 5 126.71 82.40 44.31 0.00 0.00 4.535 2 6 126.95 80.12 46.83 0.00 0.00 4.311 3 1 0.00 181.12 0.00 34.60 0.00 3 2 0.00 159.58 0.00 47.26 104.77 3 3 0.00 142.63 0.00 57.22 118, 23 3,112 3 4 0.00 128.93 0.00 65.27 128.56 2.991 3 5 0.00 117.63 0.00 71.91 136.73 2,798 3 6 0.00 108.15 0.00 77 , 48 143.35 3,271 4 1 0.00 104.33 0.00 79, 73 148.83 3.258 4 2 0.00 86.08 0.00 90, 45 156.12 3.062 4 3 0.00 73.27 0.00 97.98 160.76 2.176 4 4 0.00 63.77 0.00 103.56 164.62 2.228 4 5 0.00 56.46 0 , 00 107.86 167.88 2.407 4 6 0.00 0.00 0.00 0.00 170.67 5.222 4 6 0.00 0.00 0.00 0.00 0.00 107 TABLE 6

Library: Plate 3 (ID; 100279) Units: mg

Row Column Water B-SM-20-TeA X-Cl-3 X-Cl-2 Phosphate Binding (mmol / g) 1 1 123.95 108.47 15.49 0.00 1 2 124.34 104.88 19.47 0.00 1 3 124.70 101.51 23.19 0.00 1 4 125.04 98.36 26.66 0.00 1 5 125.36 95.40 29.97 0.00 3.958 1 6 126.66 92.61 33.06 0.00 4.309 2 1 126.13 88.30 37.82 0.00 4,417 2 2 126.47 85.14 41.33 0.00 4,424 2 3 128.78 82 , 19 44.59 0.00 4.92 2 4 127.08 79.44 47.84 0.00 4.407 2 5 127.36 76.87 50.49 0.00 4.14 2 6 127.62 74.46 53 , 16.00 0.00414 3 1 0.00 118.41 0.00 26.19 3 2 0.00 102.78 0.00 29.56 3 3 0.00 90.80 0.00 32.14 3 4 0.00 81.32 0.00 34.18 3 5 0.00 73.64 0.00 35.84 3 6 0.00 87.28 0.00 37.21 2.237 4 1 0.00 58.81 0 , 00 39.03 2.403 4 2 0.00 53.43 0.00 40.19 2.704 4 3 0.00 48.96 0.00 41.15 2.614 4 4 0.00 45.17 0.00 41.97 1,714 4 5 0.00 41.93 0.00 42.67 2.294 4 6 0.00 0.00 0.00 0.00 5,295 108 TABLE 7

Table 1 Table 1 Table 1 Table 2 Table 2 Table 2 Table 2 Table 2 Table B-SM-20-TeA B-SM-22-DA Table 1 (ID: 100353) 33.97 24.05 1 2 117.71 9.19 44.82 31.73 1 3 100.13 7.82 52.42 37.12 5.838 1 4 87.12 6.80 58.05 41.10 5 , 38 1 5 77.10 8.02 62.39 44.17 5.549 1 6 69.15 5.40 65.83 4 6.61 5.826 2 1 64.71 5.05 67.75 47.97 5.452 2 2 57.99 4.53 70.66 50.03 3,358 2 3 52.54 4.10 73.01 51.70 3, 45 2 4 48.02 3.75 74.97 53.08 4.27 2 5 44 , 22 3.45 76.61 54.24 3.469 2 6 40.98 3.20 78.02 55.24 4.058 3 1 111.71 26.16 39.87 28.23 3 2 89.37 20.93 51 , 04 38.14 3 3 74.48 17.44 58.49 41.41 5.154 3 4 63.85 14.95 63.81 45.18 5.784 3 5 55 87 87.08 67.80 48.01 5.596 3 6 4 9.66 11.63 70.91 50.20 5.287 4 1 46.24 10.83 72.62 51.42 5.261 4 2 41.13 9.63 75.17 53.23 4.743 4 37, 04 8.67 77.22 54.67 4.076 4 5 33.69 7.89 78.90 55.86 3.924 4 5 30, 90 7.24 80.29 56.85 2,898 4 6 0.00 0.00 , 00.00 5,287 TABLE 8

Library: Plate 1 (ID; 100384) Unit: mg

Row Column X-Cl-3 B-SM-22-DA Water NaOH Binding to phosphate (mmol / g) 1 1 643.88 422.44 1752.36 227.94 1 2 692.40 378.56 1743.80 245 , 12.362 1 3 731.79 342.94 1736.85 259.06 4.09 1 4 764.40 313.44 1731.10 270.61 3.186 1 5 791.85 288.62 1726.26 280.33 2.951 1 6 815.27 267.44 1722.12 288.62 2.005 2 1 64388 422.44 1752.36 227.94 2 2 692.40 378.56 1743.80 245.12 2 3 731.79 342.94 1736 , 85 259.06 2 4 764.40 313.44 1731.10 270.61 4,794 2 5 791.85 288.62 1726.26 280.33 2 6 815.27 267.44 1722.12 288.62 4.332 3 1 643.88 422.44 1752.36 227.94 3 2 692.40 378.56 1743.80 245.12 3 3 731.79 342.94 1736.85 259.06 3 4 764.40 313.44 1731 , 10 270.61 4.511 3 5 791.85 288.62 1726.26 280.33 5.086 3 6 815.27 267.44 1722.12 288.62 4.61 4 1 643.88 422.44 1752.36 227 , 94 4 2 692.40 378.56 1743.80 245.12 4 3 731.79 342.94 1736.85 259.06 4 4 764.40 313.44 1731.10 270.61 4 5 791.85 288 , 62 1726.26 280.33 4.816 4 6 0.00 0.00 0.00 0.00 5.17 1 10

Example 3: Synthesis of 1,3-diaminopropane / epichlorohydrin crosslinked beads prepared in a suspension process

A 3 liter reaction vessel was used, comprising a three-necked round bottom flask with four lateral outlets. The reaction flask was fitted with an oil heating bath, cold water reflux condenser and mechanical stirrer with a 3 inch propeller. Into this reaction vessel was introduced a solution of 1,3-diaminopropane (90.2 g, 1.21 mol) dissolved in 90.2 g of water, surfactant (branched dodecylbenzenesulfonic acid sodium salt, 6.4 g dissolved in 100 g of water) and 1 kg of toluene. This initial charge was stirred at 600 rpm for 2 minutes and then reduced to 300 rpm for 10 minutes before epichlorohydrin was added. The speed of 300 rpm was maintained for the remaining time of the experiment. The solution was warmed to 80 ° C and also maintained at this temperature throughout the experiment.

In a separate vessel a solution of epichlorohydrin in toluene 40% by mass was prepared. Using a syringe pump, 1.2 equivalents of epichlorohydrin (134.7 g, (1.45 mol)) was added to the initial charge of the reaction vessel over a period of 3 hours. The reaction was continued for a further 2 hours before 0.75 equivalents of sodium hydroxide (36.5 g (0.91 mol)) was added in a 40 wt% solution. Sodium hydroxide solution was added to the reaction through a syringe pump over a period of 2.5 hours. The reaction mixture was maintained at 80 ° C for an additional 8 hours. 111

After this period the beads were purified by removal of toluene, washing with 1000 ml of acetone, followed by methanol, 20% NaOH solution (to remove the surfactant) and then twice more with deionized water. The beads were dried by freezing for 3 days to give a fine white powder of 160 g weight (92% yield) and an average diameter of 93 μl.

Example 4: Synthesis of cross-linked 1,3-Diaminopropane / 1,3-Dichloropropane polymer

Using water as the solvent, 1000 mg of B-SM-22-DA was mixed with 1524 mg of X-Cl-3 and 2524 mg of water in a 20 ml scintillation flask. The reaction was subjected to magnetic stirring and maintained at a temperature of 80 ° C overnight, followed by a temperature of 90 ° C for a further two hours. The reaction mixture was purified to 34% by weight (1716 mg) by performing 3 washing steps in water / centrifugation to give 144.7 mg of the powder of the polymer of the present example.

Example 5: Synthesis of 1,3-Diaminopropane / 1,3-Dichloropropane Crosslinked Polymer

Using water as a solvent, 2000 mg of B-SM-22-DA was mixed with 3048 mg of X-Cl-3 and 5048 mg of water in a 20 ml scintillation vial. The reaction was subjected to magnetic stirring and kept at a temperature of 80 ° C overnight.

After 3 hours of reaction, 3597 mg of

NaOH 30% by weight in water to retain the acid produced during the reaction since the crosslinker used was an alkyl halide. The reaction mixture was purified to 20.3% by weight (2773.5 mg) by performing 3 washing steps in water / centrifugation to give 591.3 mg powder of the polymer of the present example.

Example 6: Synthesis of crosslinked beads prepared with 1,3-Diaminopropane / 1,3-dichloropropane using a prepolymer approach

Preparation of prepolymer The reaction vessel used was a 250 mL two-necked round bottom flask equipped with a reflux condenser of cold water, magnetic stirrer and under argon. A solution of 1,3-diaminopropane (31.15 g, 0.42 mol) dissolved in 30.15 g of water is introduced into the reaction vessel. This initial charge is stirred up to 300 rpm. The solution was warmed to 80 ° C and maintained at this temperature throughout the experiment. Using a syringe pump, 1 equivalent (47.47 g, 40.0 mL, 0.42 mol) of 1,3-dichloropropane (Aldrich 99%) was added over a period of 2 hours. The reaction was continued for a further 2 hours before 10 mol% (relative to 1,3-diaminopropane) of sodium hydroxide (1.68 g (0.042 mol) of NaOH was added and made to a 40% by weight in water). The sodium hydroxide solution was added to the reaction via a pipette over a period of 2 minutes. The reaction was maintained at 80 ° C for an additional 4 hours. The solution at 80 ° C is viscous and upon cooling to 25 ° C becomes a solid buffer which is rapidly soluble in water. 113

Purification

To the solid buffer is added water, washed with 200 mL of water and 200 mL of MeOH. This is then added to a 1 L beaker containing a 50/50 solution of MeOH / isopropyl alcohol. The white polymer precipitated. After placing the suspension in a centrifuge, the liquid supernatant is discarded. This process is repeated using isopropyl alcohol 2 more times. The white precipitate is then dried under reduced pressure at room temperature to remove the isopropyl alcohol. Weight of insulated polymer: Mn (GPC over a polyethyleneimine standard) - 600. Synthesis of crosslinked particles

The white prepolymer (8.7 g) was placed in a flask with 1.3 g of branched dodecylbenzenesulfonic acid sodium salt (30 wt% solution in water) and 34.8 g of toluene. This resulted in a 20 wt% solution of polymer suspended in toluene. The polymer was milled to micron range with a mechanical mill (Mark: IKA, Model: Ultra-Turax T8). 2.2 g of the resulting suspension was charged into a 10 mL reaction flask fitted with a heating mantle, mechanical stirrer and syringe pump. The reaction flask was charged with an additional 3779 mg of toluene. The flask was heated to 80 ° C and the stirrer (500 RPM) was added. After 3 hours of stirring at this temperature, epichlorohydrin (112.2 mg, 0.0012 mol) was added over a period of 1.5 hours. The reaction was allowed to proceed for an additional 2 hours before 224.4 mg (0.0056 mol) of sodium hydroxide (in a 40% solution, by weight in water) was added, which was supplied over a period of 2 hours. The reaction was allowed to cool to room temperature and stirring was stopped. The beads were purified by removing the toluene, washing with methanol and then with a 20% NaOH solution (to remove the surfactant) and twice more with deionized water. The beads were frozen for 3 days to give a fine white powder. The binding capacity measured in a non-interfering buffer was 3.85 mmol / g.

Example 7: Synthesis and isolation of a low molecular weight polymer (prepolymer) prepared with 1,3-Diaminopropane / 1,3-dichloropropane 1

Abbreviations used in the following examples:

Epichlorohydrin: ECH

N, N, Ν ', Ν'-tetrakis (3-aminopropyl) -1,4-diaminobutane: BTA BC: binding capacity

In this Example the effect of varying the ratio of monomer (in this case, a prepolymer) relative to the solvent in the reaction mixture on binding capacity and rate of expansion is demonstrated. This Example describes a process involving two parts: first, the synthesis of a soluble prepolymer addition product of 1,3-diaminopropane and 1,3-dichloropropane, and second, the preparation of insoluble beads by further crosslinking the prepolymer with ECH . The second reaction consisted of an inverse suspension process wherein the ratio of water to the prepolymer was varied. The impact of this variation on binding and dilation performance was evaluated. Prepolymer Synthesis

Step 1 (Preparation of prepolymer): The reaction vessel used was a 250 mL two-necked round bottom flask equipped with a cold water reflux condenser, magnetic stirrer and used under an argon atmosphere. In this reaction vessel was introduced a solution of 1,3-diaminopropane (31.15 g, 0.42 mol) dissolved in 30.15 g of water. This initial charge was stirred at 300 rpm. The solution was warmed to 80 ° C and maintained at this temperature throughout the experiment. Using a syringe pump, 1 equivalent (47.47 g, 40.0 mL, 0.42 mol) of 1,3-dichloropropane (Aldrich 99%) was added over a period of 2 hours. The reaction was continued for a further 2 hours before 10 mol% (relative to 1,3-diaminopropane) of sodium hydroxide (1.68 g (0.042 mol) of NaOH was added and made to a 40% by weight in water). The sodium hydroxide solution was added to the reaction via a pipette over a period of 2 minutes. The reaction was maintained at 80 ° C for an additional 4 hours. The solution at 80 ° C was viscous and upon cooling to 25 ° C became a rapidly soluble solid buffer in water.

Step 2 (Purification): Water was added to the solid buffer, washing with 200 mL of water and 200 mL of MeOH. This was then added to a 1 L beaker containing a 50/50 solution of MeOH / isopropyl alcohol. The white polymer precipitated. After centrifugation, the liquid supernatant was discarded. This process was repeated using isopropyl alcohol 2 more times. The white precipitate was then dried under reduced pressure at room temperature to remove the isopropyl alcohol. Molecular weight of the isolated polymer: Mn (GPC vs. polyethyleneimine standard) ~ 600. Synthesis of micrometric range-cross-linked particles prepared with 1,3-Diaminopropane / 1,3-dichloropropane prepolymer in a parallel polymerization reactor of 24 wells semicontinuous.

The white prepolymer 1 (8.7 g) was placed in a flask with 1.3 g of branched dodecylbenzenesulfonic acid sodium salt (30 wt% solution in water) and 34.8 g of toluene. This yielded a 20 wt% solution of polymer suspended in toluene. The emulsion was triturated to droplets in the micrometric range with a high shear stress homogenizer (Brand: IKA, Model: Ultra-Turax T8). 2.2 g of the resulting emulsion was charged to 24 of the 10 mL reaction flasks of the reactor which was equipped with a heating mantle, a mechanical stirrer and a syringe pump. Another 3779 mg of toluene was charged to each reaction flask. The flasks were heated to 80 ° C and the stirrer (500 RPM) was attached. Water was added to the tubes in an amount necessary to produce various prepolymer / water ratios. After 3 hours of stirring at this temperature, the desired amount of epichlorohydrin was added (in this Example, epichlorohydrin was added to an amount equal to 20% of the dry weight of the prepolymer) over a period of 1.5 hours. The reaction was allowed to proceed for a further 2 hours before 224.4 mg (0.0056 mol) of sodium hydroxide (in a 40 wt% solution in water) was added, which was provided over a period of 2 hours. The reaction was allowed to cool to room temperature and stirring was stopped. The beads were purified by removing the toluene, washing with methanol and then with a 20% NaOH solution (to remove the surfactant) and then with HCl to protonate the bead. The beads were then washed twice with deionized water to remove excess HCI. The beads were freeze-dried for 3 days to give a fine white powder.

The polymer beads thus obtained were analyzed for binding capacity (BC) in non-interfering buffer and in a GI mock, and in relation to the rate of dilation. The results are summarized in Table 9. TABLE 9 1,3-Diamino-propane / 1,3-dichloropropane-1,3-diene beads. Effect of ratio of monomer to water on Binding Capacity and Dilatation Proportion BC (mmol / g) non BC (mmol / g) Dilatation (g of monomer / mock interfering water GI H20 / g polymer) 1.67 3.85 1.54 2.92 1.42 3.68 1.43 3.34 1.25 3.61 1.34 3.50 1.11 3.55 1.34 3.70 0.83 3.31 1, 16 5.22 0.55 2.90 0.91 14.00

These results show that the binding capacities in both non-interfering buffer and GI mock increase as the ratio of monomer to water increases, while the rate of expansion decreases and reaches the desired range. 118

Example 8: Synthesis of cross-linked particles in the micrometric range from crude BTA / ECH gel crushed using a 24-well parallel polymerization reactor

In this Example the effect of varying the amount of crosslinker relative to the monomer in the binding capacity and the rate of expansion is demonstrated.

The following stock solution was prepared: 2 molar equivalents of concentrated HCl was added to 1 molar equivalent of BTA over a period of 2 hours. Water was then added to the solution so that the resulting solution reached the following composition in% by weight: 45 wt% BTA, 10 wt% HCl, 45 wt% water. Into each flask of a 24-well reactor using 5 ml flasks was placed 0.6 g of the prepared stock solution. The desired amount of epichlorohydrin was added to each flask to give the monomer: crosslinker ratio to be tested. The reactor was heated to 80 ° C for 9 hours. The reactor was allowed to cool. To each flask was added water to dilate the resulting polymer. The gel was then crushed to obtain particles in the micrometer range with a high shear strength homogenizer (Brand: IKA, Model: Ultra-Turax T8). The particles were then purged from the water, washed with methanol and then with a 20% NaOH solution and then with HCl to protonate the amine functionalized particle. The particles were then washed twice with deionized water to remove excess HCl. The particles were dried by freezing for 3 days to give a fine white powder.

The results of binding and dilatation studies are summarized in Table 10. TABLE 10

BTA / ECH Gel: Binding and Dilation Capability Data against the crosslinking content. Raw gels (the monomer: water ratio is 75% by weight of Bow-Tie (2HCl) in water). The proportions of monomer: water vary from 3.5 (ECH: BTA = 0.85) to 4.8 (EHC: BTA = 6.4). ECH: BTA BC (mmol / g) g) Simulation GI Dilatation (g Water / g Polymer) 0.70 0.00 0.00 0.00 0.85 2.23 0.35 1.00 2.46 0.49 16.68 1.15 2.57 0.49 10.98 1.30 2.84 0.58 6.17 1.45 2.91 0.65 4.69 1.60 2.91 0.77 3.85 1.79 2.88 0, 85 3.13 1.98 0.00 0, 98 2.7 / 2.00 2.46 1.00 2.55 2.00 2.46 1.00 2.55 2.16 2.73 0.99 2.46 2.35 2.67 0.96 2.20 2.40 2.17 0.93 1.97 2.40 2.17 0.93 1.97 2.80 1.86 0.82 1, 81 2.80 1.86 0.82 r ^ r 3.20 1.63 0.73 1.84 3.20 1.63 0.73 1.84 3.60 1.28 0.64 1.57 3 , 60 1.28 0, 64 1.57 4.00 1.09 0.58 1.57 4.00 1.09 0.58 1.57 4.40 0.88 0.45 2.03 4.40 0.88 0.45 2.03 4.90 0.42 0.35 1.47 4.90 0.42 0.35 1.47 5.40 0.42 0.28 1.5U 120 (continued)

BTA / ECH Gel: Binding and Dilation Capability Data against the crosslinking content. Raw gels (the monomer: water ratio is 75% by weight of Bow-Tie (2HCl) in water). The proportions of monomer: water vary from 3.5 (ECH: BTA = 0.85) to 4.8 (EHC: BTA = 6.4). ECH: BTA BC (mmol / g) g) Simulation GI Dilatation (g Water / g Polymer) 5, 40 0.42 0.28 1.50 5, 90 0.07 0.27 1.55 5, 90 0.07 0.27 1.55 6, 40 0.06 0.22 1.55 6, 40 0.06 0.22 1.55

These data show that the binding capacity in the GI simulacrum passes through a maximum as the ratio of crosslinker to the amine varies. In this particular system the optimum binding capacity in the GI simulacrum is observed at a crosslinker ratio of 1.8 to 2.8, corresponding to an NC value of 3.6 to 5.6 respectively. In this range of crosslinking the rate of dilation is minimal. Similar tests may be performed for other monomers and crosslinkers using this polymerization protocol to determine the ratio that gives the desired results for the particular use in which the polymer will be placed.

Example 9: Synthesis of ΒΤΆ / ECH beads crosslinked in the micrometric range via reverse suspension

The following stock solution was prepared: 2 molar equivalents of concentrated HCl was added to 1 molar equivalent of BTA over a period of 2 hours. Water and surfactant (branched dodecylbenzenesulfonic acid sodium salt, 30% by weight in water) were then added to the solution so that the resulting solution reached the following compositions by weight: BTA 41.8% by weight, HCl 9 , 4% by weight, water 41.1% by weight, surfactant (30% by weight in water), 7.7% by weight. The reaction vessel used was a 0.25-liter three-necked round bottom flask, with four side outlets, provided with an oil heating bath, cold water reflux condenser and mechanical stirrer with a 1-inch propeller. 25 g of the prepared stock solution and 75 g of toluene were charged to this reaction vessel.

In a separate vessel was prepared a 40% by mass solution of epichlorohydrin in toluene. Using a syringe pump, the desired amount of ECH was added over a period of 90 minutes. The reaction was continued for another 2 hours before starting dehydration using Dean Stark equipment. The end of the reaction was reached when all the water had been eliminated from the system. The beads were purified by removing the toluene, washing with methanol and then with a 20% NaOH solution (to remove the surfactant) and then with HCl to protonate the bead. The beads were then washed twice with deionized water to remove excess HCl. The beads were lyophilized for 3 days to give a fine white powder.

The results of the binding and dilation studies are summarized in Table 11. TABLE 11 BTA / ECH Gel Pearls: Bonding and Dilatation Capacities against Crosslinking Content. Molar ratio ECH: BTA BC (mmol / g) Non interfering BC (mmol / g) Digested Meal Dilatation (g Water / g polymer) 1.00 2.50 0.58 25.29 1.00 2.77 0 , 55 13.01 1.25 2.97 0.65 7.69 1.25 3.03 0.61 7.07 1.50 3.13 0.71 4.41 1.50 3.14 0.69 3.99 1.75 3.13 0.78 3.06 1.75 3.10 0.87 3.41 2.00 3.07 0.99 3.13 2.00 2.80 1.00 2, 82 2.00 2.82 0.73 3.17 2.50 2.76 1.03 2.48 3.00 2.56 0.82 2.40 3.50 0.00 0.71 2.28 3 , 00 2.32 0.70 2.25 3.00 2.61 0.80 2.03 3.50 2.81 0.59 1.85 4.00 0.00 0.58 1.99 4.00 2.19 0.77 1.93 4.50 2.11 0.30 1.99 5.00 1.96 0.55 1.72

These results show that the binding capacity in the GI simulacrum passes through a maximum as the ratio of crosslinker to the amine is varied. In this particular system the optimum binding capacity in the GI simulacrum is observed at a crosslinker ratio of 1.75 to 3, corresponding to an NC value of 3.5 to 6 respectively. 123

Within this range of crosslinking the rate of expansion is minimal. Similar tests may be performed for other monomers and crosslinkers using this polymerization protocol to determine the ratio that gives the desired results for the particular use in which the polymer will be placed.

Example 10: Synthesis of cross-linked particles in the micrometric range from crude polyallylamine / ECH crude gel using a 24-well parallel polymerization reactor

This Example illustrates the synthesis of a polymer using a high molecular weight monomer and varying the proportions of monomer relative to water in the reaction mixture. The conditions used were identical to those described in Example 8, except that polyallylamine (MW = 60,000 g / mole) was used in place of BTA. The ratio of ECH to the allylamine-like structural unit was 1: 0.106 (corresponding to a CN of 2.2). The initial ratio of polyallylamine to water was changed from 1: 1 to 1.4. As a comparative example, cross-linked polyallylamine of Renagel tablets was used.

Molar ratio BC (mmol / g) BC (mmol / g) Dilatation rate (g of Amine in Non-Meal Water / g of Polymer) relative to digested interfering water o, 3.66 0.92 19.00 0, 33 4.12 1.36 6.00 0.50 4.20 1.62 4.00 Renagel 3.85 1.40 9.00 124

These data indicate that a higher proportion of amine relative to water led to a lower rate of dilation and was accompanied by greater binding capacity in the GI simulacrum. Similar tests may be performed for other monomers and crosslinkers using this polymerization protocol to determine the ratio that gives the desired results for the particular use in which the polymer will be placed.

Example 11: Measurement of interference at the connection level

This example illustrates the measurement of binding interference using a polymer of the invention and, for comparison, a polymer of the prior art. A crosslinked polyamine (EC172A) material was prepared according to the protocol described in Example 4, having a BTA: ECH molar ratio of 2.5, and a ratio of (BTA + ECH) to water of 1.73. Interference on binding was compared to Renagel. 0 &quot; degree of interference on binding &quot; or &quot; interference in connection, &quot; as used herein refers to a fractional decrease in the binding capacity of the target ion observed between a binding experiment in a non-interfering buffer and a gastrointestinal (GI) mock at the same concentration of target anion at equilibrium. A binding isotherm was first obtained in a non-interfering buffer representing the binding capacity against equilibrium phosphate concentration for a variety of phosphate concentrations. This isotherm was then adjusted by an exponential function to predict the binding capacity at any phosphate concentration. The binding capacity measured in the GI simulacrum was then determined in the same isotherm, representing the point of the 125 phosphate concentration against the phosphate binding at equilibrium for the simulacrum 01 and extending the vertical line to its point of interception with the non-interfering isotherm . The degree of interference was then calculated as (BCNI-BCGI) / BCNI * 100. Interference on binding to EC127A is shown in the following Table and in Figure 3.

(MmHg) BC (mmol / g) BC Predicted (mmol / g) Interference (%) 6.25 3.31 1.18 2.17 45.7 6.25 3.28 1.19 2 , 16 45.0 6.25 3.24 1.21 2.15 44.0 Interference on binding to RENAGEL is shown in the following Table and in Figure 4.

Mean (mM) Peq (mM) BC (mmol / g) Expected BC (mmol / g) Interference (%) 6.25 2.70 1.42 4.53 66.7 6.25 2.54 1.48 4 , 46 66.7 Interference on binding to EG127A is about 34% lower than that of RENAGEL. 126

Example 12: Ion binding properties in ex vivo liquids collected by aspiration in humans

A crosslinked polyamine (EC172A) material was prepared according to the protocol described in Example 4, having a BTA: ECH molar ratio of 2.5, and a ratio (BTA + ECH) relative to water of 1.73. The material was then tested for phosphate binding in an aspirated human liquid beaker as described in Example 1. The EC172A phosphate binding was compared to the crosslinked polyallylamine active drug isolated from Renagel (Genzyme). The EC172A exhibits a much lower level of interference, as well as a much lower rate of dilation (2.5 vs. 9 for the Renagel)

Mean Mean DP Mean BC BC DP% predicted% of (mM) (mmol / g) (mmol / g) (mmol / g) interference Renagel API 2.37 0.01 1.32 0.00 4.37 70 EC172A 1.55 0.04 1.64 0.02 1.68 2.5

In another experiment, both materials, EC172A and Renagel, were used in a human ex-vivo liquid harvested by different aspiration to quantify the degree of interference in the phosphate binding produced by competing solutes such as citrate anions and bile acids. The citrate and bile acid anions were metered by ion chromatography and enzyme assay respectively. The data presented below (mean of six volunteers) indicate that the polymer of the present invention exhibits much better selectivity and overall phosphate binding. 127 (P04) BC (P04) [citrate] BC (citrate) (Biliary Acids) BC (Biliares) mM mmol / g mM mmol / g mM mmol / g Control (without polymer) 5,722 1,667 4,928 Renagel 3,019 1,078 0,596 0,429 1, 32 1.443 EC172A 1.78 1.573 1.316 0.141 4.65 0.109

Example 13: Measurement of gel porosity using the solute partitioning technique

This Example illustrates the measurement of the porosity of the gel. Measurements were made on a polymer of the invention and on a commercially available phosphate-binding polymer for comparison. As a polymer of the invention, a crosslinked polyamine (EC172A) material was prepared according to the protocol described in Example 10, having a BTA: ECH molar ratio of 2.5, and a ratio of (BTA + BCH) relative to the water of 1.73. For comparison, the same porosity measurements were performed on Renagel.

The probes were 8 polyethylene glycols (PEG) with a MW of 200 to 20,000 Da and 4 poly (ethylene oxides) (PEO) (30,000 to 23,000 Da).

All probes were dissolved in 30 mM ammonium acetate buffer pH 5.5 (5 g / L concentration). The solutions of the probes were added to the washing of EC172A with HCl (5 mL / g) and washing of Renagel with HCl (15 mL / g dry gel) pre-weighed; it was then stirred for 4 days in a Vortexer. 128

The solutions of the probes were diluted 10x prior to LC analysis using a Polymer Lab Light Scattering Evaporative Detector (to work in the linear range of the detector and ensure that the ratio of peak areas is equal to the concentration ratio by weight) . The calculation of the volume Not accessible = msw + [l-cantes / Cdepois] / where msw is the amount of water collected by the gel (g / g dry gel), msoiv the amount of water, in which the probe at the beginning [g / g of dry gel] Cantes and CdePois: concentrations of the probe before and after equilibration The Cantes / Cdepois ratio is equal to the proportion of the areas of the peaks obtained by LC.

The results of this Comparative Example are shown in Figures 5 and 6; Figure 5 shows the results in terms of molecular weight while Figure 6 illustrates the results in terms of the size of the solutes. EC127A shows a constant molecular exclusion for solutes with higher PMs up to 200, compared to Renagel, which has a decreasing PM exclusion as high as 1000.

Example 14: Post-modification of beads with chloropropylamine hydrochloride

Preparation of the stock solution: • Chloropropylamine hydrochloride (B-SM-34-A) in water at 50% by weight - d = 1,132 • Sodium hydroxide in water at 30% by weight (by dilution of a 50% by weight) - d = 1.335 129 Synthesis:

FR-0005-144, a phosphate binder polymer prepared according to Example 9, having a BTArECH molar ratio of 2.5, and a (BTA + ECH): water ratio of 1.73, was used as a substrate for further amination: The FR-0005-144 beads were transferred into 4ml flasks (two 4x6 plates each containing 21 flasks) and water, chloropropylamine hydrochloride stock solution and sodium hydroxide stock solution was added using a robot supplier of liquids. The vials were sealed with a capsule and the plates were placed in reactors provided with an individual heating and stirring system.

Heating and stirring was continued for 12 hours: The reactor temperature was adjusted to 85øC and the stirring speed was set at 200 rpm.

Purification:

Each of the disposable culture tube materials (16x100mm) was washed and washed once with methanol, twice with 1M hydrochloric acid solution in water, and three times with water. Each time the beads were separated by centrifugation.

It was then dried in a lyophilizer and analyzed for the Digested Meal, Non-interfering Buffer and Expansion Rate. The results are shown below in Table 12 and Fig.

Characteristics of polymers prepared by post-modifying pearls with chloropropylamine hydrochloride FR-0005-144 aCfUci B-SM-3 4-A NaOH B-SM-34-A weight ratio ¥ .s. (Rrmol / g) BC non-interfering (rrmol / g) Dilatation rate (g water / g gel) 222 Molar ratio of NaOH (vs. B-SM- , 1 864.5 22.2 1.71 0.1 0.25 0.94 2.84 2.91 233.3 883.0 46.7 3.59 0.2 0.25 0.91 2.94 2.69 203, 7 749, 0 61.1 4.70 0.3 0.25 0.95 2.85 2.83 209, 1 746.3 83.6, 6.43, 0.25, 0.25, 97 2.91 2.64 209 723.5 104.5 8.04 0.5 0.25 0.97 2.89 2.58 - - - - - 227 761.3 136.2 10, 48 0.6 0.25 0.96 2.90 2.60 235 762.8 164.5 12.65 65 0.7 0.25 1.00 2.97 2.67 231.3 725.9 9.08 14.23 0.8 0.25 0.99 2.88 2.86 278.5844, 1 250.7 19.28 0.9 0.25 0.99 2.90 3.38 236.2 690, 4 236.2 18.17 1.0 0.25 1.00 2.96 2.73 't 204.1 792.9 9 20.4' λ 1 4 , 92 2.81 2.85 271 1021.5 54.2 8.34 0.2 0, 5 0.95 2.81 2.74 247 902.5 74.1 11.40 0.3 0.05 , 97 2.85 2.85 225, 5 797, 9 90.2 13.87 0.4 0, 5 0.97 2, 93 2.61 238.2 815, 4 119.1 18.32 0.5 0.51 2.84 2.68 270.77 ft 0.89 2.73 2.98 199.76 60, 5 119.8 18.43 0.6 0, 5 0.98 2.91 2.70 230, 6 736, 161.4 24.83 0.7 0.5, 1.01, 3.02, 46 221.3 680, 9 177.0 27.23 0.8 0, 5 0.98 2, 92 2.58 212, 5 629, 3 191.3 29, 42 0.90, 1.02 , 04 2.61 200, 4 570, 4 200.4 30.83 1.0 0, 5 1.06 2. 93 2.46 0 0 0 0 213, 1,826.17 21.3 4, 1 0.75 0.94 2.80 2.92 203.7 764.66 40.7 9.40 0.2 0.75 0.94 2.81 2.82 212.4 771.18 63.7 14 , 70 0.3 0.75 0.97 2.84 3.04 218.2 765.38 87.3 20.14 0.4 0.75 1.00 2.88 2.99 203.4688.43 101.7 23.47 0.5 0.75 1.03 2.90 2.64 * 214.3 CJW / J «- / 128, b 29, b7 U / c U / /« ./ 1 / U./2.94 2.5u 131 (continued) FR-0005- 144 water B-SM-3 4-A NaOH B-SM-34-A weight ratio vs (Mmol / g) BC non-interfering (mmol / g) Dilatation rate (g water / g gel) 228 FR-0005-144 NaOH molar ratio (vs. B-SM- , 8 718.09 160.2 36, 95 0.7 0.75 1.04 2.95 2.60 235.2 709.23 188.2 43.41 0.8 0.75 1.08 3.02 2.55 216.8 627.06 195.1 45.02 0.9 0.75 1.00 2.95 2.65 206.7 572.41 206.7 47.69 1.0 0.75 1, 00 3.03 2.48% v: 1: 1: 1: 1: 1, n, 199.7 772.69 20.0 6.14 0.1 1.0 0.97 2.75 2 , 85 206.4 771.62 41.3 12.70 0.2 1.0 0.97 2.77 3.30 216 779.26 64.8 19.94 0.3 1.0 0.98 2, 83 2.93 213.3 741.63 85.3 26.25 0.4 1.0 1.00 2.85 3.43 212.9127 4 106.5 32.75 0.5 1.0 1 , 04 2.95 2.66 193.3 0, 0.95 2.73 2.95 240.66 773.63 144.4 44.41 0.6 1.0 1.02 2.94 2.88 294 , 5 908.43 206.2 63.42 0.7 1.0 1.07 2.94 2.58 214.1 632.43 171.3 52.69 0.8 1.0 1.06 3.05 2.60 205, 5 580.15 185, 0 56, 90 0.9 1.0 1.08 3.04 2.66 201.2 541, 7 201.20 61, 90 1.0 1.0 1, 09 3.07 2.91. ,

Example 15: Synthesis of cross-linked particles in the phosphate printing micrometer range from Ν, Ν, Ν ', Ν' - (tetra-3-aminopropyl) -1,4-diaminobutane / epichlorohydrin

The following mother solution was prepared: 1 molar equivalent of Phosphoric Acid (Aldrich, 85% in water) was added to 1 molar equivalent of Ν, Ν, Ν ', N' - (tetra-3-aminopropyl) -1 , 4-diaminobutane over a period of 2 hours. Water was then added to the solution so that the resulting solution reached the following composition in weight%: ΝΝ, Ν ', N' - (tetra-3-aminopropyl) -1,4-diaminobutane 42% by weight, H 3 PO 4 13% by weight, water 45% by weight. The reactor contained 24 wells of 5 mL flasks, each flask containing a magnetic stir bar. At each flask was placed 0.6-0.7 g of the prepared stock solution. The stirrers were switched on. The desired amount of pure epichlorohydrin was added to each vial. The reactor was heated to 60 ° C for 1 hour and then warmed to 80 ° C for 8 hours. The reactor was allowed to cool. To each flask was added water to dilate the resulting gel. The gel was transferred to a 4x6 plate with 10 mL test tubes. The gel was then crushed to micron size particles with a mechanical grinder (Mark: IKA, Model: Ultra-Turax T8). The particles were purified by removing the water, washing with methanol and further washing with a 20% NaOH solution. The gel particles were subsequently washed with 1.0 mol HCl, mixed for 30 minutes, the gel was then allowed to stand and the supernatant was decanted. This process was repeated 5 times to protonate the amine-functionalized particle with Chloride and replace the bound H3PO4. The gel particles were then washed with a 20% NaOH solution to deprotonate the amine functionalized gel particles. The gel particles were then washed twice with deionized water to remove excess NaOH / NaCl. The gel particles were lyophilized for 3 days to give a fine white powder. The synthesis is summarized in Table 13. TABLE 13 Synthesis of gels which were Molecularly impregnated with phosphoric acid. (Mg) B-SM-20-TeA (moles) phosphoric acid (mg) water (mg) X-EP-1 (mg) X-EP-1 (moles) B 20 TeA / H3P04 X-EP-1 / B-SM-20- TeA Gel present in the well 1.0 1.0 347.5 0.0011 107.7 369.8 71.1 0.0008 1.000 0.70 X 1.0 2.0 339.5 0.0011 105.2 361.4 79.4 0.0009 1.00 0.00 0.80 X 1.0 3.0 337.7 0.0011 104.6 359.4 88.8 0.0010 1.00 0.90 / 1.0 4.0 352.1 0.0011 109.1 374.8 102.9 0.0011 1.00 1.00 1.0 5, 0 355.4 0.0011 110.1 378.2 114.3 0.0012 1.00 1.10 ✓ 1.0 6, 0 366.1 0.0012 113.4 389.6 128 , 4 0.0014 1.00 1.20 ✓ 2.0 1.0 355.3 0.0011 110.1 378.1 135.0 0.0015 1.00 1.30 2.0 2.338, 6 0.0011 104.9 360.4 138.6 0.0015 1.00 1.40 / 2.0 3, 0 356.2 0.0011 110.4 379.1 156.2 0.0017 1.00 1.50 2,0 2.0 4.0 349, 7 0.0011 108.3 372.2 163.5 0.0018 0.99 1.61 / 2.0 5, 0 342.2 0.0011 106.0 364.2 170.0 0.0018 1.00 1.70 ✓ 2.0 6, 0 351, 4 0.0011 108, 9 374.1 184.9 0.0020 1.00 1.80 ✓ 3.0 1.0 364.1 0.0012 112.8 387.5 212.8 0.0023 1.00 2.00 3.0 2, 0 351.2 0.001 1 108.8 373.8 246.4 0.0027 1.00 2.40 ✓ 3.0 3, 0 358.3 0.0011 111.0 381.4 293.2 0.0032 1.00 2.81 ✓ 3.0 4.0 340.2 0.0011 105.4 362.1 318.2 0.0034 1.00 3.20 ✓ 3.0 5, 0 368, 9 0.0012 114.3 392.6 388.2 0.0042 1.00 1.00 3.59 / 3.0 6, 0 360, 5 0.0011 111.7 383.7 421.5 0.0046 1.00 / 4.0 345.3 0.0011 107.0 367.5 444.0 0.0048 1.00 1.00 4.40 / 4.0 2, 0 364, 0 0.0012 112.8 387.4 510.7 0.0055 1 , 00 4.80 / 4.0 3, 0 351.2 0.0011 108.8 373.7 533.7 0.0058 1.00, 5.20 / 4.0 4.0 365, 5 0.0012 113 , 2 389.0 598.3 0.0065 0.99 5, 63 / 4.0 5, 0 358, 5 0.0011 111.1 381.6 628.8 0.0068 1.00,

The polymers synthesized as described bind phosphate.

While preferred embodiments of the present inventive concept have been disclosed and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. A great number of variations, alterations and substitutions will come to the minds of those skilled in the art without departing from the inventive concept. It will be understood that various alternatives may be applied to the embodiments of the inventive concept described herein in the practice of the inventive concept. The following claims define the invention.

Lisbon, August 24, 2007 135

Claims (20)

A cross-linked amine polymer comprising an amine monomer of formula wherein n is 3, 4 or 5, and the amine is crosslinked with a crosslinking agent. The crosslinked amine polymer of claim 1, wherein the amine monomer is Ν, Ν, Ν '' '' - tetrakis (3-aminopropyl) -1,4-diaminobutane. The polymer of any one of the preceding claims, wherein the crosslinking agent has at least two functional groups. The polymer of claim 3, wherein the crosslinking agent is selected from the group consisting of 1,3-dichloropropane and epichlorohydrin. The polymer of any one of the preceding claims, comprising epichlorohydrin-crosslinked Ν, Ν, Ν ', N'-tetrakis (3-aminopropyl) -1,4-diaminobutane, wherein the polymer is produced by a process in that the ratio of the initial concentration of Ν, Ν, Ν ', N'-tetrakis (3-aminopropyl) -1,4-diaminobutane to water is from about 1: 3 to 4: 1. The polymer of any of claims 1 to 4, wherein the polymer is a phosphate-binding polymer comprising the cross-linked N, N, Ν ', Ν'-tetrakis (3-aminopropyl) -1,4-diaminobutane amine monomer with epichlorohydrin, and the polymer is produced by a process in which the total epichlorohydrin crosslinker added to the reaction mixture ranges from about 200% to about 300 mol% of the total amine monomer content N, N, , N'-tetrakis (3-aminopropyl) -1,4-diaminobutane in the reaction mixture. The polymer of claim 6, wherein the polymer is in the form of spherical beads. The polymer of any one of the preceding claims, wherein the crosslinking reaction is performed in crude solution or dispersed medium. The polymer of any one of the preceding claims, wherein the crosslinking reaction leading to the formation of the gel is carried out using either a) homogeneous process or ii) heterogeneous process. The polymer of any one of the preceding claims, wherein the polymer is a phosphate-binding polymer 2 characterized by a dilation rate, measured in isotonic medium at neutral pH, of less than 5. The polymer of any one of the preceding claims, wherein the polymer binds the phosphate ion in vivo with a binding activity greater than 0.5 mmol / g. The polymer of any one of the preceding claims, wherein the polymer is formulated as a free, non-contracted amine. The polymer of any one of the preceding claims, wherein the polymer has a transition temperature greater than about 30 ° C. A pharmaceutical composition comprising a polymer of any one of claims 1 to 13 and a pharmaceutically acceptable excipient. The pharmaceutical composition of claim 14, wherein the pharmaceutical composition is formulated with a chewable tablet or a liquid formulation. Use of a polymer of any one of claims 1 to 13 in the preparation of a medicament. Use of a polymer of any one of claims 1 to 13 in the preparation of a medicament for treating a condition in which there is an excess ion. Use of a polymer of any one of claims 1 to 13 in the preparation of a medicament for removing an anion from an animal by administering an effective amount of the polymer to the animal. The use of any one of claims 16 to 18 wherein the polymer is used in the preparation of a medicament for treating a selected pathology of hyperphosphataemia, hypocalcemia, hyperparathyroidism, depressed renal synthesis of calcitriol, tetany due to hypocalcemia, renal failure, ectopic calcification in soft tissues, chronic renal failure and anabolic metabolism. The non-therapeutic use of a polymer of any one of claims 1 to 13. Lisbon, 24 AUGUST 2007 4